ES2373834B1 - DERIVATIVES OF DIFLUOROBENCYL ETHANOLAMINES WITH ANTIMICROBIAL ACTIVITY. - Google Patents

Abstract

Derivatives of difluorobenzyl ethanolamines with antimicrobial activity, belongs to the field of medical chemistry. It refers to a method of synthesis of fluorinated ethanolamines molecules, specifically difluorobenzyl ethanolamines, as well as to these new molecules and their growth inhibitory activity of species of the Mycobacteríum and Nocardía genera, pathogenic bacteria that are involved in the development of diseases such as tuberculosis or leprosy, as well as other infections at the lung, cutaneous or central nervous system level.

Description

Derivatives of di-fluorobenzyl ethanolamines with antimicrobial activity.

Technical sector of the invention

The present invention belongs to the field of medical chemistry. It refers to a method of synthesis of fluorinated ethanolamines molecules, specifically di-fluorobenzyl ethanolamines, as well as said new molecules and their growth inhibitory activity of species of the Mycobacterium and Nocardia genera, pathogenic bacteria that are involved in the development of diseases such as tuberculosis or leprosy, as well as other infections at the lung, cutaneous or central nervous system level.

Background of the invention

The search for new therapeutic agents is a priority in the fight for the control of infectious diseases.1 Antimicrobials constitute the fundamental basis for the treatment of infectious diseases, such as those caused by Mycobacterium, specifically Mycobacterium tuberculosis that is the cause of tuberculosis human and responsible for almost three million deaths per year in the world, being also one of the opportunistic pathogens with the highest incidence in patients with HIV +. Studies carried out in recent years have identified the molecular targets of current drugs in use and their most frequent resistance mechanisms.2 Some of the active compounds against M. tuberculosis (TB) are isoniazid and pyracinamide, which affect the synthesis of fatty acids by the bacteria, or ethambutol that is involved in the biosynthesis processes of the bacterial cell wall.

The new anti-TB agents must act on a target that is essential for the survival of mycobacteria and must be active throughout their growth cycle, both inside and outside human cells during infection. The M. tuberculosis cell envelope is responsible for its relatively small size compared to other bacteria, has hydrophobic properties and is refractory3, so it resists the action of a drug without changing state or destroying itself. Structurally it consists of a peptidoglycan (PG) layer that contains alternating units of N-acetylglucosamine and N-glycolylmuramic acid. The tetrapeptide side chains are linked in carbon number six of the murmic acid residue which in turn is linked via phosphodiester with arabinogalactan (AG). AG is a polymer composed of residues of D-galactofuranosa and D-arabinofuranosa, it is extremely rare in nature, but necessary for the viability of mycobacteria. On the other hand, it is bound to mycolic acids of 70-90 carbon atoms by esterification of the terminal residues of arabinose. PG, AG and mycolic acids are the three components that make up the cell wall nucleus of the micolyl-arabinogalactan-peptidoglycan (mAGP) complex. The mAGP contains a lipoarabinomannan (LAM) framework that is involved in the virulence and immuno-pathogenesis of TB. It contains a large number of polar and apolar lipids on the outside of the cell wall that are associated and distributed over the cell surface. It is very similar to the cell membrane of gram-negative bacteria. Due to its great complexity and low permeability it contributes significantly to resistance against many therapeutic agents.

Ethambutol, a synthetic amino alcohol that has activity against many strains of the Mycobacterium genus, interrupts the biosynthesis of arabinano that is essential for the formation of AG and LAM, inhibiting the arabinosyl transferase enzyme, 4 interferes with the conversion of D-glucose to D -Arabinous for the transformation of arabinose to AG. Arabinosyl transferase is responsible for the formation of the mAGP complex and if its biosynthetic pathway is inhibited, permeability in the cell wall is increased. An increase in permeability increases, and facilitates the passage of molecules into the cytoplasmic membrane. The design of substrates capable of behaving in this way would represent an important step in the new generation of antibacterials.

In recent decades, some drugs have been approved for the treatment of tuberculosis, and others are in clinical phase, 5 which shows the urgent need for new therapies to combat this disease.

In the state of the art there are several documents that describe obtaining tri-fluoromethylamines. Thus, Kuduk et al, 6 "Asymmetric addition reactions of Grignard reagents to chiral 2-tri fl uoromethyl tert-butyl (Ellman) sulfinimine-ethanol adducts" Tetrahedron Letters 2006, 47, 2377-2381, describe that the addition of Grignard reagents to Tri fl uoromethyl tert-butyl sulphimimine-ethanol adducts allow tri-fluoromethylamines to be protected with high yield and a diastereoselectivity between good and excellent. However, the result of stereochemical addition is the opposite of what was expected by controlled chelation the transition state.

Philippe, Christine et al.7 “Synthesis of New Tri fl uoromethylated Hydroxyethylamine-Based Scaffolds” Eur. J. Org. Chem. 2009, 5215-5223, refers to the synthesis of new tri-fluoromethyl-hydroexyethylamines by means of epoxides whose ring opening they obtain with amino compounds, including aliphatic and other amines. The results on the comparison of water and fluorinated and non-fluorinated alcohols in their use as solvents are also given. Regioselectivity is observed and the stereochemistry of the compounds is maintained. Specifically, they describe a process for obtaining tri-fluoromethyl hydroxyethylamine derivatives from epoxides using nitrogen compounds.

Dos Santos, Mickael et al. 8 “Improved Ritter reaction with CF3-containing oxirane for an Access to central units of protease inhibitors” Tetrahedron Letters 2009, 50 857-85, investigates the influence of alcohols fluorinated on BF3.Et2O in reactions Ritter type. The tri-fluoroethanol / BF3.Et2O system allows access to the central unit of protease inhibitors through the opening of the fluorinated epoxide cycle with several nitriles.

However, there is still a need to develop new drugs functionally different from those currently described. This search for new active substrates has included both natural products and products of synthetic origin. No one is oblivious to the fact that a problem associated with the discovery of a new drug is the need to study large libraries of compounds using procedures with good resolution.

Object of the invention

Therefore, the present invention relates to the synthesis of new di-fluorobenzyl ethanolamines represented by the general formula (I) of Figure 1, as well as their pharmacological application as antimicrobials,

where:

a) the protecting group R may be benzyloxycarbonyl, tert-butyloxycarbonyl; Y

b) Ar are derivatives of benzene, furan, thiophene or pyridine, which may independently possess fluorine, chlorine, bromine, iodine and methoxy substituents.

Detailed description of the invention

The replacement of hydrogen atoms with fluorine atoms in the design of new active agents is a general strategy in the design of new therapeutic agents9. Specifically, the design of new agents with antimicrobial activity should have as main objective the synthesis of active substrates with optimal permeability and metabolic stability.

Therefore, the present invention relates to the obtaining and biological behavior of di-fluorobenzyl ethanolamines that has not been explored in the state of the art since the di-fluorobenzyl grouping represents a metabolically stable system that can improve the pharmacodynamics of potentially active compounds10. Regarding the method of obtaining, it has been observed that the epoxidation process used in the present invention leads to the formation of the two possible diastereoisomers, which, once formed, can be separated by chromatography, which has allowed the generation of the four possible diastereoisomers, using the two previously generated allylamines and reducing the total number of stages of the synthesis. In addition, because the double bond of alpha-fl uorinated allylamines is strongly deactivated, the use of oxone11 (potassium peroximonosulfate) as an oxidant has allowed the first example of direct epoxidation thereof to be carried out, which is not possible with other conventional oxidants. Comparing with the procedures described in the works of Philippe et al.7 it should be noted that they use a different synthetic methodology and that they only investigate the preparation of one of the diastereoisomers while in the present invention using oxone, the two epoxides have been achieved .

Taking these considerations into account, a library of di-fluorobenzyl ethanolamines of formula (I) has been designed following the general strategy shown in Scheme 1. The ethanolamine cluster, as shown, could be prepared by regioselective opening of the epoxy ring of a fluorinated epoxyamime II by reaction with benzyl amines. This epoxide, which contains all the necessary carbon skeleton, could be obtained by stereoselective epoxidation of fluorinated allylamines III conveniently functionalized. Taking into account the possibility of generating allylamines in chiral form, the subsequent epoxidation allows all possible isomers to be prepared.

Scheme 1

Taking into account that no examples of synthesis of di-fluorobenzyl ethanolamines have been included in the literature, and that the presence of CF2 groups in benzyl positions improves the metabolic stability of the substrates so that the pharmacodynamics of potentially active compounds could be improved, the investigation It was initiated by the preparation of the library shown in Figure 2.

Synthesis of the di-fluorobenzyl ethanolamines of the invention

Fluorinated imines are fundamental and interesting symptoms for the introduction of nitrogen and fluorine atoms into polyfunctionalized structures.12,13 In fact, tri-fluoromethylimines have previously been used in the synthesis of protease inhibitors.14 In the present invention, it is used as key step the convenient functionalization of allylamine 4 prepared by stereoselective alkylation of the fluorobenzyl imines 5.

Although it was initially considered to exploit the tandem protocol for the synthesis of N-PMP allylamine described by the authors of the present application previously15, which uses a chiral equivalent of vinyl anion, the strategy did not provide the expected results in preliminary trials. As an alternative, it was studied to apply the diastereoselective synthesis methodology described by Kuduk et al 6, based on the use of Ellman's chiral N-tert-butylsulphimimines to prepare chiral amines.14 This procedure avoids the isolation of fl uorinated N-tert-butylsulphimines, 15 which are very reactive substrates, and has allowed the preparation of chiral di-fluorobenzyl allylamines with good yields and selectivity, as shown in Scheme 2 below.

Scheme 2

The precursor necessary for the synthesis of imines is not commercial, so it was necessary to synthesize it from ethyl benzoylformate 6. The instability of ethyl hemiacetals 7, make it necessary to directly transform it into hemiaminals. Reaction with (R) -terc-butanosulfinamide in the presence of TiOEt4 at 70 ° C, overnight, provided hemiamine 8, as a 3: 1 mixture of the two possible diastereoisomers, (53% overall yield for the four-step sequence from 6). The unequivocal separation and allocation of the stereochemistry of both epimers was irrelevant, since both lead to the corresponding di-fluorobenzyl allylamine 4 with 76% yield when treated with two equivalents of low magnesium vinyl bromide in THF.18 The compound 4 'was prepared using the same procedure and with similar yields using in this case the (S) -terc-butanosulfinamide, as shown in Scheme 3 below.

Scheme 3

This strategy provides the chiral di fl uorobenzyl allylamines 4 and 4 ′ key for the present invention, with a high enantiomeric purity, 20: 1 d.r., as deduced from the 19F nuclear magnetic resonance spectrum analysis. The absolute stereochemistry was proposed assuming the model proposed by Kuduk et al.6, although this end was subsequently confirmed by determining the X-ray structure of brosylate crystal 9 obtained from allylamine 4, as shown in the Scheme 4 and Figure 2.

The next necessary step was the hydrolysis of the chiral auxiliary due to its incompatibility under epoxidation conditions. Hydrolysis using hydrochloric acid (4 M) in dioxane, 19 followed by the reaction of the resulting ammonium salt with base and capture of the free amine with benzyloxycarbonyl chloride provided carbamate 10 with a 90% yield, while when the Reaction was carried out with Boc2O, compound 11 was obtained in 82% yield (Scheme 5). Attempts to alternatively also introduce an acetate group were unsuccessful due to its lability in the chromatographic puri fi cation process.

Scheme 5

Fortunately, epoxidation of 10-fluorobenzyl allylamine with tri-fluoromethylmethyldioxyran in heterogeneous medium 20 gave a 74% yield of an equimolecular mixture (deduced from its 1 H NMR) from the two isomeric epoxides 12a and 12b which were separated by column chromatography on silica gel. The application of the same reaction conditions on carbamate 11 yielded the mixture of epoxides 13a and 13b although with a somewhat lower yield. It should be noted at this point that both mixtures showed some instability in the column chromatography process on silica gel, being more pronounced in the case of the N-Boc carbamate.

With the intention of establishing the stereochemistry of the new stereogenic centers created, the epimers 12a and 12b were converted independently into their corresponding oxazolidinones 14, by treatment in a first stage with dibenzylamine in isopropanol at 70 ° C, followed by reaction with sodium hydride in tetrahydrofuran at 0 ° C (Scheme 6). Stereochemistry was determined by careful spectroscopic analysis using different NMR techniques, particularly by correlations observed in the heteronuclear nOe (HOESY) 1H-19F experiments. Oxazolidininone 14 shows a spatial proximity between the CF2 group and the H5 atom, therefore, the assigned stereochemistry is 4R and 5S. While for oxazolidinone 15, crossing peaks between the CF2 group and the methylene group are observed, which is in accordance with the stereochemistry shown, 4R and 5R, subsequently confirmed by an X-ray diffraction study thereof, Figure 3 .

Following the same sequence of reactions applied to the 4 'enantiomeric fl uorobenzyl allylamine, the derived N-Cbz and N-Boc carbamates, compounds 10' and 11 'respectively, were obtained, which subsequently were transformed into the four epoxides 12'a, 12'b , 13'a and 13'b with the yields indicated in Scheme 7.

Scheme 7

To achieve the proposed objective, it was only necessary to carry out the coupling of the epoxides with the benzylamines to transform those into the objective β-amino alcohols of the work. With the intention of optimizing the process, different opening reaction conditions of epoxide 13b with 3-iodobenzylamine were studied. The best results were when the epoxide was heated to isopropanol reflux in the presence of the amine to drive 93% yield to the di-fluorobenzyl amino alcohol 16b. Similarly, epoxide 13a was converted into the di-fluorobenzyl amino alcohol 16a in 87% yield, according to Scheme 8.

Scheme 8

The application of the same conditions to the epoxides 12, led to the four possible opening isomers 17 with the yields shown in Scheme 9.

Scheme 9

Finally, the synthesis work was completed following the same protocol as before with the preparation of the chemo library of the di-fluorobenzyl ethanolamines shown in Table 1, with the indicated yields. The structure of the products obtained are shown below Table 1.

ES 2 373 834 A1

TABLE 1

Structures products table 1

As additional information for a possible study of the relationship between structure and reactivity, other di-fluorobenzyl ethanolamines (27a and 28b) were prepared in order to study on the one hand the influence of the absence of the iodine atom and on the other hand the absence of the amine free NH group.

Scheme 10

Experimental part

Synthesis of the compounds

All melting points were measured on a hot platform Ko fl er apparatus. The optical rotations were determined by a 5 cm long cuvette and the values of [α] D are expressed in units of 10-1 deg cm2 g − 1. High resolution mass spectra have been obtained by electronic impact (EI) at 70 eV, by rapid atom bombardment (FAB) or by electrospray ionization (ESI). The 1H NMR spectra were performed in CDCl3 at 300 or 400 MHz, and those of 13C NMR at 75 or 100 MHz. The 1H spectra were referenced to the residual CHCl3 (δ 7.26), the 13C spectra to the central component of the triplet of CDCl3 at δ 77.0, and the 19F spectra to the internal reference (CCl3F). The degrees of carbon substitution were established by DEPT pulse sequences. A combination of COZY, HMQC and NOE experiments were used when necessary for the assignment of chemical shifts of 1H and 13C.

Column chromatography refers to fl ash chromatography and was carried out on silica gel Merck 60, of grain 230-400. All operations involving air sensitive reagents were performed under an inert atmosphere of dry nitrogen or argon, using syringes, oven-dried glassware and freshly dried and distilled solvents.

Ethyl 2,2-di-fluoro-2-phenylacetate. To a solution of ethyl benzoyl formate (2.25 g, 12.0 mmol) dissolved in 8 mL of dry dichloromethane, in a 0 ° C bath, it was slowly added with a 50% fl uor addition funnel in toluene (24 mmol). A catalytic amount of absolute ethanol was then added and the reaction mixture was left under continuous stirring at room temperature for 20 h. After confirming by final layer chromatography the complete disappearance of the starting substrate, hydrolysis was carried out on a water / ice mixture and extraction with dichloromethane. The combined organic phases were dried over anhydrous Na2SO4 and concentrated under reduced pressure. Purification of the crude by fl ash chromatography [n-hexane: AcOEt (6: 1)] allowed to obtain the product as a transparent liquid, with a yield of 99% (2.4 g). 1H-NMR (300 MHz, CDCl3) δ 1.30 (t, J = 7.1 Hz, 3H), 4.30 (c, J =

7.1 Hz, 2H), 7.42-7.53 (m, 3H), 7.60-7.63 (m, 2H). 13C-NMR (75.5 MHz, CDCl3) δ 12.7, 62.0, 112.4 (t, JCF = 251.7 Hz), 124.4 (t, JCF = 6.0 Hz), 127.6, 129.9, 131.8 (t, JCF = 25.6 Hz), 163.2 t , JCF = 35.4 Hz). 19F-NMR (282.4 MHz, CDCl3) δ -104.4 (s, 2F). EMAR (EI +) calculated for C10H10O2F2 [M +]: 219.0262, found: 219.0263.

2,2-Di-fluoro-2-phenylethanol. To a suspension of LiAlH4 (319 mg, 8.4 mmol) in THF (26 mL) at 0 ° C was added, dropwise with an addition funnel, a solution of ethyl 2-phenyl-2,2-di-fluoroacetate ( 560 mg, 2.8 mmol) in 30 mL of THF. It was then left under stirring until the disappearance of the starting substrate by CCF. The reaction was then treated with Na2SO4 · H2O, the mixture obtained was filtered, washing repeatedly with dichloromethane, and the filtrate was concentrated. The resulting clear oil was purified by fl ash chromatography [n-Hexane: AcOEt (4: 1)], providing the alcohol (438 mg, 96%). 1H-NMR (CDCl3, 300 MHz) δ 2.23 (sa, 1H), 3.96 (t, JHF = 13.5 Hz, 2H), 7.43-7.48 (m, 3H), 7.48-7.54 (m, 2H). 13C-NMR (CDCl3, 75.5 MHz) δ 66.0 (t, JCF = 32.3 Hz), 120.6 (t, JCF = 243.6 Hz), 125.4 (t, JCF = 6.3 Hz), 128.5, 130.3 (t, JCF = 1.3 Hz ), 134.3 (t, JCF = 25.4 Hz). 19F-NMR (CDCl3, 282.4 MHz) δ

107.7 (t, JHF = 13.5 Hz, 2F). EMAR (EI +) calculated for C8H8F2O [M +]: 158.0543, found: 158.0542.

N - [(1RS) -1-ethoxy-2-phenyl-2,2-di fl uoroethyl] (R) -t-butylsul fi namide (8). A solution of DMSO (19.6 mmol) in dichloromethane (6 mL) was added to a solution of oxalyl chloride (9.4 mmol) in 6 mL of dichloromethane at -60 ° C. The mixture was allowed to stir at the same temperature for 5 minutes. After this time, a solution of 2-phenyl-2,2-di-fluoroethanol (1.23 g, 7.8 mmol) in 21 mL of dichloromethane was added dropwise. The resulting mixture was stirred at -60 ° C for 15 minutes. After that time, a solution of triethylamine (39.3 mmol) in 6 mL of dichloromethane was added. After stirring at the same temperature for another 5 minutes, it was allowed to temper for two hours until reaching room temperature. When the starting substrate was consumed, 15 mL of ethanol was added and left under constant stirring at room temperature for 3 hours. The reaction mixture was concentrated to dryness, obtaining a whitish solid residue that was resuspended in ethyl ether and filtered. The ethereal filtrate was concentrated again under reduced pressure, obtaining a light yellow oil that was used immediately in the next reaction step. In a sealed tube under inert conditions, 1-ethoxy-2-phenyl-2,2-di-fluoroethanol (1.0 mmol), (R) -t-butylsulphamamide (1.0 mmol) and titanium tetraethoxide [Ti (OEt) 4] (5.0 mmol) were added ). The reaction mixture was allowed to stir at 70 ° C overnight. After that time it was diluted with 5 mL of ethyl acetate, treated with ice-water and the resulting mixture was extracted with ethyl acetate. The combined organic phases were washed with brine and dried over anhydrous Na2SO4. The solvent was evaporated under reduced pressure and the yellow oil obtained by 1 H NMR indicated that it was a mixture of the diastereoisomers in a 3: 1 ratio. An analytical amount was purified by fl ash chromatography, using n-hexane: AcOEt (2: 1) as eluent (161 mg, 53%, 2 steps).

Major Diastereoisomer: Yellow solid, mp: 52-54 ° C (CH2Cl2). [α] D25 = -59.14 (c 1.0, CHCl3). 1H-NMR (CDCl3, 300 MHz) δ 1.15 (t, J = 6.9 Hz, 3H), 1.18 (s, 9H), 3.52 (dc, J = 9.3, 6.9 Hz, 1H), 3.54 (d, J = 10.2 Hz, 1H),

3.96 (dc, J = 9.3, 6.9 Hz, 1H), 4.72 (ddd, J = 10.2, 8.5, 4.2 Hz, 1H), 7.40-7.44 (m, 3H), 7.49-7.50 (m, 2H). 13C-NMR (CDCl3.75MHz) δ 14.6, 22.4, 56.8, 65.0, 89.1 (dd, JCF = 35.2 Hz, JCF = 32.5 Hz), 118.7 (t, JCF = 248.4 Hz), 126.3 (t, JCF = 6.2 Hz ), 128.0, 130.2, 133.1 (t, JCF = 25.4 Hz). 19F-NMR (CDCl3, 282 MHz) δ -111.3 (dd, JFF = 249.0 Hz, JHF = 8.5 Hz, 1F), -104.2 (dd, JFF = 249.0, JHF = 4.2 Hz, 1F). EMAR (FAB) calculated for C14H22F2NO2S [M + H +]: 306.1339, found: 306.1349.

Minority diastereoisomer: Yellow solid, mp: 45-47 ° C (CH2Cl2). [α] D25 = -12.05 (c 1.0, CHCl3). 1H-NMR (CDCl3, 300 MHz) δ 1.09 (t, J = 6.9 Hz, 3H), 1.23 (s, 9H), 3.40 (dc, J = 9.3, 6.9 Hz, 1H), 3.76 (dc, J = 9.3 , 6.9 Hz, 1H), 4.05 (d, J = 9.3 Hz, 1H), 4.77 (ddd, J = 10.4, 9.3, 3.1 Hz, 1H), 7.42-7.44 (m, 3H), 7.55-7.57 (m, 2H). 13C-NMR (CDCl3.75MHz) δ 14.6, 22.5, 57.1, 64.9. 87.7 (dd, JCF = 36.7 Hz, JCF = 32.2 Hz), 118.6 (t, JCF = 249.2 Hz), 126.5 (t, JCF = 6.4 Hz), 128.1, 130.2, 133.0 (t, JCF = 25.4 Hz). 19F-NMR (CDCl3, 282 MHz) δ -113.0 (dd, JFF = 251.9 Hz, JHF = 10.4 Hz, 1F), -103.0 (dd, JFF = 249.0, JHF = 3.1 Hz, 1F). EMAR (FAB) calculated for C14H22F2NO2S [M + H +]: 306.1339, found: 306.1335.

N - [(1SR) -1-Ethoxy-2-phenyl-2,2-di fl uoroethyl] (S) -t-butylsul fi namide (8 ’). When (S) -t-butylsulfinamide was used, similar results were obtained. 1.9 g of alcohol led to a mixture of both hemiaminals which, after chromatography, provided 1.0 g (27%) of the majority diastereoisomer [α] D25 = +71.07 (c 1.0, CHCl3), and 351 mg (10% ) of the minor diastereoisomer: [α] D25 = +16.85 (c 1.0, CHCl3).

[(R) -t-Butyl sulfyl] [(1R) -1- (phenyldi fl uoromethyl) allyl] amine (4). To a solution of the mixture of hemiamine 8 (2.66 g, 8.7 mmol) in dichloromethane (95 mL) at -60 ° C was added a 1 M solution in THF of vinyl magnesium bromide (21.7 mL). The mixture was allowed to slowly warm to -25 ° C, when the starting substrate had completely disappeared. At this point, the mixture was hydrolyzed with aqueous NH4Cl, allowed to warm to room temperature and extracted with dichloromethane. The combined organic phases were washed with brine and dried over anhydrous Na2SO4. Finally, the solvent was removed under reduced pressure and the residue obtained was chromatographed on SiO2 to provide sulfinamide 4 as a yellowish oil, in a 20: 1 diastereomeric ratio, as determined by 1 H-NMR and 19 F-NMR. [α] D25 = -101.49 (c 1.0, CHCl3). 1H-NMR (CDCl3, 300 MHz) δ 1.19 (s, 9H), 3.57 (d, J = 3.0 Hz, 1H), 4.27-4.38 (m, 1H), 5.32 (d, J = 16.8 Hz, 1H), 5.36 (d, J = 9.9 Hz, 1H), 5.62 (ddd, J = 16.8, 9.9, 7.7 Hz, 1H), 7.40.7.47 (m, 5H). 13C-NMR (CDCl3, 75.5 MHz) δ 22.4, 56.0, 63.4 (t, JCF = 28.9 Hz), 120.9 (t, JCF =

249.5 Hz), 122.9, 126.1 (t, JCF = 6.3 Hz), 128.3, 130.3 (t, JCF = 4.0 Hz), 130.4 (t, JCF = 1.7 Hz). 133.7 (t; JCF = 26.4 Hz). 19F-NMR (CDCl3, 282.4 MHz) δ -107.3 (dd, JFF = 244.3 Hz, JHF = 13.0 Hz, 1F), -103.4 (dd, JFF = 244.3 Hz, JHF = 7.9 Hz, 1F). EMAR (FAB) calculated for C14H20F2NOS [M + H +]: 288.1234, found: 288.1235.

(S) -t-Butyl sulphyl] [(1S) -1- (phenyldi fl uoromethyl) allyl] amine (4 ’). When 1.35 g of the 8 'hemiamine mixture was used, 1.05 g of allylamine (85%) were obtained after chromatography. [α] D25 = +105.20 (c 1.0, CHCl3).

Benzyloxycarbonyl [(1R) -1- (phenyldi fl uoromethyl) allyl] amine (10). To a solution of allylamine 4 (200 mg, 0.7 mmol) in anhydrous methanol (4.5 mL) was added 1.8 mL of a solution of HCl (4 M) in dioxane, and allowed to stir at room temperature for 2 hours After this time, the reaction crude was concentrated to dryness and the white residue obtained was redissolved in 4.5 mL of dioxane. Then, K2CO3 (2.1 mmol), benzoyl chloride (3.5 mmol) and a catalytic amount of dimethylaminopyridine (DMAP) (0.1 mmol) were successively added at room temperature. The reaction mixture was left with continuous stirring at room temperature for 18 hours. After this period, the resulting mixture was hydrolyzed with brine, the two phases were separated and the aqueous phase was extracted with AcOEt. The combined organic phases were washed with brine and dried over Na2SO4. After evaporating the solvent under reduced pressure, the oil obtained was subjected to fl ash chromatography, using nhexane: ethyl ether (4: 1) as eluent, obtaining a transparent oil corresponding to allylamine 10, with a 91% yield. Mp: 64-66 ° C (CH2Cl2). [α] D25 = +21.14 (c 1.0, CHCl3). 1 H-NMR (CDCl 3, 300 MHz) δ 4.83-5.00

(m, 1H), 5.08 (s, 1H), 5.08 (s, 2H), 5.29 (d, J = 12.0 Hz, 1H), 5.30 (d, J = 16.2 Hz, 1H), 5.84 (ddd, J = 16.2, 10.2, 5.4 Hz, 1H), 7.30-7.37 (m, 5H), 7.37-7.50 (m, 5H). 13C-NMR (CDCl3, 75.5 MHz) δ58.5 (t, JCF = 29.3 Hz), 67.1, 119.2, 120.7 (t, JCF = 258.4 Hz), 125.7 (t, JCF = 6.3 Hz), 128.0 , 128.2, 128.3, 128.5, 130.2 (t, JCF = 1.4 Hz), 130.6 (t, JCF = 2.6 Hz),

134.1 (t, JCF = 25.6 Hz), 136.0, 155.6. 19F-NMR (CDCl3, 282.4 MHz) δ-106.9 (dd, 2JFF = 247.2 Hz, JHF = 12.7 Hz, 1F), -104.5 (dd, JFF = 247.7 Hz, JHF = 11.9 Hz, 1F). EMAR (EI +) calculated for C18H17F2NO2 [M +]: 317.1227, found: 317.1229.

Benzyloxycarbonyl [(1R) -1- (phenyldi fl uoromethyl) allyl] amine (10 ’). 1.3 g of alkylamine 4 'provided 970 mg of alkylamine N-Cbz 10' (68%). [α] D25 = -9.18 (c 1.0, CHCl3).

(R) -1-Phenyl-1,1-di-fluoro-3-butenyl-2-carbamate tert-butyl (11). To a solution of allylamine 4 (1.0 g, 3.5 mmol) in anhydrous methanol (21.6 mL) 8.7 mL of a solution of HCl (4 M) in dioxane was added, and allowed to stir at room temperature for 2 hours. After this time, the reaction crude was concentrated to dryness and the white residue obtained was redissolved in 21.6 mL of dioxane. Then, K2CO3 (10.4 mmol), di-t-butyl dicarbonate (3.5 mmol) and a catalytic amount of dimethylaminopyridine (DMAP) (0.1 mmol) were successively added at room temperature. The reaction mixture was left with continuous stirring at room temperature for 52 hours. After this period, the resulting mixture was hydrolyzed with brine, the two phases were separated and the aqueous phase was extracted with AcOEt. The combined organic phases were washed with brine and dried over Na2SO4. After evaporating the solvent under reduced pressure, the oil obtained was subjected to fl ash chromatography, using n-hexane: ethyl ether (7: 1) as eluent, obtaining a white solid corresponding to allylamine 11 (805 mg, 82%). Mp: 49-51 ° C (CH2Cl2). [α] D25 = +23.67 (c 1.0, CHCl3). 1H-NMR (CDCl3, 300 MHz) δ1.37 (s, 9H), 4.80-4.85 (m, 1H), 4.82 (sa, 1H), 5.28 (d, J = 9.3 Hz, 1H), 5.29 (d, J = 18.0 Hz, 1H), 5.80-5.89 (m, 1H), 7.40-7.43 (m, 3H), 7.47

7.50 (m, 2H). 13C-NMR (CDCl3, 75.5 MHz) δ27.3, 57.8 (t, JCF = 28.1 Hz), 85.1, 118.8, 120.9 (t, JCF = 249.4 Hz), 125.7 (t, JCF = 6.2 Hz), 128, 130.0 , 130.9 (t, JCF = 2.9 Hz), 134.4 (t, JCF = 25.7 Hz), 146.7. 19F-NMR (CDCl3, 282.4 MHz) δ -106.5 (dd, JFF = 251.6 Hz, JHF = 10.4 Hz, 1F), -105.3 (dd, JFF = 251.6 Hz, JHF = 11.3 Hz, 1F). EMAR (ESI +) calculated for C15H19F2NNaO2 [M + Na +]: 306.1282, found: 306.1284.

(S) -1-Phenyl-1,1-di-fluoro-3-butenyl-2-carbamate tert-butyl (11 ’). 1.7 g of allylamine 4 'provided 430 mg (41%) of N-Boc-allylamine 11'. [α] D25 = -20.78 (c 1.0, CHCl3).

(R) -2-Phenyl-2,2-di-fluoro-1 - [(R) oxyran-2-yl] ethyl benzyl ethylcarbamate (12a) and (R) -2-phenyl-2,2-di-fluoro-1- [ (S) benzyl oxyran2-yl] ethylcarbamate (12b). To a solution of allylamine 10 (951 mg, 3.0 mmol) in acetonitrile (26 mL) was added 13 mL of an aqueous solution of Na2EDTA · 2H2O (4 · 10-4 M). The solution was cooled to 0 ° C and tri-fluoromethylmethyl ketone (1.0 mL) was added. Subsequently, on the reaction mixture at 0 ° C, NaHCO3 (17.8 mmol) and Oxone (5.7 mmol) were added together in a single portion. After 3 h of reaction, the suspension was filtered, the phases were separated and the aqueous phase was extracted with dichloromethane. After removing the solvent in vacuo, the solid residue obtained was subjected to fl ash chromatography, using n-hexane: ethyl ether as eluent with a polarity gradient (10: 1 to 6: 1), which allowed to obtain 441 mg ( 44%) of 12a and 308 mg (31%) of 12b.

Isomer R, R (12a). White solid, mp: 68-70 ° C (CH2Cl2). [α] D25 = +4.38 (c 1.0, CHCl3). 1H-NMR (CDCl3, 300 MHz) δ2.36 (dd, J = 4.5, 2.4 Hz, 1H), 2.61 (t, J = 4.5 Hz, 1H), 3.18-3.22 (m, 1H), 4.52 (td, J = 12.0, 9Hz, 1H), 4.97 (d, J = 9.0 Hz, 1H), 5.03 (s, 2H), 7.24-7.54 (m, 10H). 13C-NMR (CDCl3, 75.5 MHz) δδ42.5, 48.0 (t, JCF = 3.6 Hz),

55.0 (t, JCF = 29.3 Hz), 67.3, 121.1 (t, JCF = 249.2 Hz), 125.6 (t, JCF = 6.4 Hz), 127.9, 128.2, 128.5, 128.5, 130.4, 133.9 (t, JCF = 25.9 Hz ), 135.9, 156.0. 19F-NMR (CDCl3, 282.4 MHz) δ-104.9 (d, JHF = 12.0 Hz, 2F). EMAR (EI +) calculated for C18H17F2NO3 [M +]: 333.1177, found: 333.1182.

Isomer R, S (12b). White solid, mp: 92-94 ° C (CH2Cl2). [α] D25 = -8.05 (c 1.0, CHCl3). 1H-NMR (CDCl3, 300 MHz) δ 2.65-2.76 (m, 2H), 3.11-3.16 (m, 1H), 4.27-4.37 (m, 1H), 4.98-5.15 (m, 1H), 5.06 (s, 2H), 7.26-7.56 (m, 10H). 13C-NMR (CDCl3, 75.5 MHz) δ 43.9, 48.9 (t, JCF = 3.4 Hz), 57.8 (t, JCF = 29.3 Hz), 67.4, 120.9 (t, JCF = 247.6 Hz), 125.4 (t, JCF = 6.3 Hz), 128.0, 128.3, 128.6, 128.6, 130.6, 133.8 (t, JCF = 25.9 Hz), 135.8, 155.7. 19F-NMR (CDCl3,

282.4 MHz) δ -106.8 (dd, JFF = 251.7 Hz, JHF = 12.5 Hz, 1F), -104.9 (d, JFF = 251.7 Hz, JHF = 13.4 Hz, 1F). EMAR (EI +) calculated for C18H17F2NO3 [M +]: 333.1177, found: 333.1175.

(S) -2-Phenyl-2,2-di-fluoro-1 - [(S) oxiran-2-yl] ethyl benzylcarbamate (12'a) and (S) -2-phenyl-2,2-di-fluoro-1 - [(R) oxyran2-yl] benzyl ethylcarbamate (12'b). 965 mg of 10 'allylamine led to a mixture of both diastereoisomeric epoxides which, after fl ash chromatography provided 307 mg (30%) of the isomer (S, S) (12'a) [α] D25 = -4.98 (c 1.0, CHCl3 ), and 441 mg (44%) of the isomer (S, R) (12'b). [α] D25 = +13.04 (c 1.0, CHCl3).

(R) -2-Phenyl-2,2-di-fluoro-1 - [(R) oxyran-2-yl] tert-butyl ethylcarbamate (13a) and (R) -2-phenyl-2,2-di-fluoro-1 - [(S) Oxyran-2-yl] tert-butyl ethylcarbamate (13b). To a solution of allylamine 11 (631 mg, 2.23 mmol) in acetonitrile (17 mL) was added 8.4 mL of an aqueous solution of Na2EDTA · 2H2O (4 · 10-4 M). The solution was cooled to 0 ° C and tri-fluoromethylmethyl ketone (4.5 mL) was added. Subsequently, on the reaction mixture at 0 ° C, NaHCO3 (35.6 mmol) and Oxone (11.1 mmol) were added together in a single portion. After 3 h of reaction, the suspension was filtered, the phases were separated and the aqueous phase was extracted with dichloromethane. The combined organic phases were washed with brine and dried over Na2SO4. anhydrous. After removing the solvent in vacuo, the solid residue obtained was subjected to fl ash chromatography, using n-hexane: ethyl ether as eluent with a polarity gradient (10: 1 to 6: 1), which allowed to obtain 187 mg ( 28%) of 13a and 277 mg (42%) of 13b.

Isomer R, R (13a). White solid, mp: 65-67 ° C (CH2Cl2). [α] D25 = +8.16 (c 1.0, CHCl3). 1H-NMR (CDCl3, 300 MHz) δ 1.32 (s, 9H), 2.51 (dd, J = 4.5, 2.4 Hz, 1H), 2.70 (dd, J = 4.5, 4.5 Hz, 1H), 3.28-3.29 (m , 1H), 4.78 (d, J = 10.2 Hz, 1H), 7.42-7.45 (m, 3H), 7.50-7.53 (m, 2H). 13C-NMR (CDCl3, 75.5 MHz) δ 28.0, 42.4, 48.0 (t, J = 3.2 Hz), 54.3 (t, J = 29.1 Hz), 80.2, 121.2 (t, J = 249.3 Hz), 125.6 (t, J = 6.3 Hz), 128.4, 130.2, 134.2 (t, J = 25.4 Hz), 155.1. 19F-NMR (CDCl3, 282.4 MHz) δ -106.0 (dd, JFF = 250.5 Hz, JHF = 11.9 Hz, 1F), -104.4 (dd, JFF = 250.5 Hz, JHF = 11.6 Hz, 1F). EMAR (FAB) calculated for C15H20F2NO3 [M + H +]: 300.1411, found: 300.1402.

Isomer R, S (13b). White solid, mp: 112-114 ° C (CH2Cl2). [α] D25 = -5.6 (c 1.0, CHCl3). 1H-NMR (300 MHz, CDCl3) δ 1.35 (s, 9H), 2.67 (dd, J = 4.5, 2.1 Hz, 1H), 2.73 (dd, J = 4.5, 4.5 Hz, 1H), 3.10-3.20 (m , 1H), 4.85 (d, J = 10.2 Hz, 1H), 7.43-7.45 (m, 3H), 7.50-7.53 (m, 2H). 13C-NMR (75.5 MHz, CDCl3) δ 28.0, 43.8, 49.0 (t, J = 3.2 Hz), 57.1 (t, J = 25.6 Hz), 80.4, 121.0 (t, J = 247.9 Hz), 125.5 (t, J = 6.3 Hz), 128.4, 130.3, 134.1 (t, J = 25.4 Hz), 154.8. 19F-NMR (CDCl3, 282.4 MHz) δ -106.4 (dd, JFF = 251.6 Hz, JHF = 11.6 Hz, 1F), -104.8 (dd, JFF = 251.6 Hz, JHF = 13.8 Hz, 1F). EMAR (FAB) calculated for C15H20F2NO3 [M + H +]: 300.1411, found: 300.1407.

(S) -2-Phenyl-2,2-di-fluoro-1 - [(S) oxyran-2-yl] tert-butyl ethylcarbamate (13'a) and (S) -2-phenyl-2,2-di-fluoro -1 - [(R) oxyran-2-yl] tert-butyl ethylcarbamate (13'b). 325 mg of 11 'allylamine led to a mixture of both diastereoisomeric epoxides which, after fl ash chromatography, provided 35 mg (10%) of the isomer (S, S) (13'a) [α] D25 = -7.32 (c 1.0, CHCl3) and 67 mg (20%) of the isomer (S, R) (13'b) [α] D25 = +6.13 (c 1.0, CHCl3).

(4R, 5S) -5 - ((Dibenzylamino) methyl) -4- (phenyl (di-fluoro) methyl) -2-oxazolidinone (14). On a solution of epoxide 12a (25 mg, 0.075 mmol) in anhydrous isopropanol (1 mL), and at room temperature, dibenzylamine (0.375 mmol) was added. The resulting mixture was heated in a sealed tube with constant stirring at 70 ° C for 5 hours. After removing the solvent under reduced pressure, the crude was purified by fl ash n-hexane: AcOEt chromatography (2: 1) on silica gel to remove excess amine. The crude obtained was dissolved in THF (1 mL) and treated at 0 ° C with NaH (pre-washed with hexane to remove the mineral oil and dried under vacuum) (0.75 mmol). The resulting mixture was allowed to stir at room temperature for 12 hours. After that time, it was hydrolyzed with water, extracted with dichloromethane and the organic phases were washed with brine and then dried over anhydrous Na2SO4. The solvent was evaporated in vacuo and the crude obtained was purified by fl ash chromatography, using n-hexane: ethyl ether (10: 1), as eluent, oxazolidinone 14 (25.2 mg, 80%) being isolated as a white solid, mp: 132-135 ° C (CH2Cl2). [α] D25 = -11.08 (c 1.0, CHCl3). 1H-NMR (300 MHz, CDCl3) δ 2.60 (dd, J = 14.0, 4.6 Hz, 1H), 2.72 (dd, J = 14.0, 6.3 Hz, 1H), 3.57 (s, 4H), 3.72 (td, J = 11.3, 3.6 Hz, 1H), 4.62 (ddd, J = 6.3, 4.6, 3.6 Hz, 1H), 5.50 (sa, 1H), 7.23-7-35 (m, 12H), 7.38-7.49 (m, 3H ). 13C-NMR (75.5 MHz, CDCl3) δ 55.1, 59.4, 60.1 (t, JCF = 31.8 Hz), 75.0, 119.7 (t, JCF = 247.6 Hz), 125.5 (t, JCF = 6.3 Hz), 127.3, 128.4, 128.8, 129.2, 130.8, 132.5 (t, JCF = 25.6 Hz), 138.6, 158.0. 19F-NMR (CDCl3, 282.4 MHz) δ -109.8 (dd, JHF = 10.9, 16.1 Hz, 2F). EMAR (FAB) calculated for C25H24F2N2O2 [M + H +]: 423.1884, found: 423.1887.

(4R, 5R) -5 - ((Dibenzylamino) methyl) -4- (phenyl (di-fluoro) methyl) -2-oxazolidinone (15). Following the same procedure, epoxy 12b led to oxazolidinone 15 with 80%, as a white solid, mp: 146-148 ° C (CH2Cl2). [α] D25 = -4.48 (c 1.0, CHCl3) .1H-NMR (300 MHz, CDCl3) δ 2.94 (d, J = 14.7 Hz, 1H), 3.01 (dd, J = 14.7, 9.3 Hz, 1H),

3.51 (d, J = 13.8 Hz, 2H), 3.76 (d, J = 13.8 Hz, 2H), 3.95 (dt, J = 18.9, 7.5 Hz, 1H), 4.81 (ddd, J = 18.9, 7.5, 2.4 Hz , 1H), 5.08 (sa, 1H) 7.13-7-43 (m, 15H). 13C-NMR (75.5 MHz, CDCl3) δ 52.0 (t, JCF = 3.5 Hz), 53.1, 58.7, 78.9, 120.2 (t, JCF = 250.3 Hz), 125.3 (t, JCF = 6.3 Hz), 127.0, 128.2, 128.8, 129.0, 131.0, 133.2 (t, JCF = 25.6 Hz), 139.0, 158.1. 19F-NMR (CDCl3, 282.4 MHz) δ -108.2 (d, JFF = 251.3 Hz, 1F), -102.3 (d, JFF = 251.3 Hz, 1F). EMAR (EI +) calculated for C25H24F2N2O2 [M +]: 422.1805, found: 422.1817.

(2R, 3S) -1-Phenyl-1,1-di-fluoro-3-hydroxy-4- (3-iodobenzylamino) butaneyl-2-carbamate benzyl (17a). To a solution of the epoxide 12a (40 mg, 0.12 mmol) in anhydrous isopropanol (0.5 mL) at room temperature 3yodobenzylamine (0.18 mmol) was added and the mixture was heated in a sealed tube at 70 ° C for 15 hours , with constant agitation, until the disappearance of the starting epoxide by CCF was observed. Then, the solvent was removed under reduced pressure and the crude obtained was purified by fl ash chromatography, using n-hexane: AcOEt (2: 1) as eluent, the amino alcohol 17a being isolated as a yellowish solid (55 mg, 81%). Mp: 85-87 ° C (CH2Cl2). [α] D25 = -20.82 (c 1.0, CHCl3). 1H-NMR (CDCl3, 300 MHz) δ 2.34 (sa, 2H), 2.58 (dd, J = 12.3, 9.3 Hz, 1H), 2.68 (dd, J = 12.3, 4.5 Hz, 1H), 3.67 (s, 2H ), 4.01 (dd, J = 9.3, 4.3 Hz, 1H), 4.18 (td, J = 13.7, 10.3 Hz, 1H), 5.04 (d, J = 12.3 Hz, 1H), 5.09 (d, J = 12.3 Hz , 1H), 5.59 (d, J = 10.3 Hz, 1H), 7.03 (t, J = 7.7 Hz, 1H), 7.19 (d, J = 7.7 Hz, 1H), 7.26-7.60 (m, 13H). 13C-NMR (CDCl3, 75.5 MHz) δ 51.4, 52.6, 57.2 (t, JCF = 28.0 Hz), 65.7, 67.1, 94.5, 121.7 (t, JCF = 250.9 Hz), 125.6 (t, JCF =

6.4 Hz), 127.2, 127.9, 128.2, 128.4, 128.5, 128.5, 130.2, 134.6 (t, JCF = 25.0 Hz), 136.2, 136.3, 137.0, 141.9, 156.3. 19F-NMR (CDCl3, 282.4 MHz) δ -104.7 (dd, JFF = 249.1 Hz, JHF = 14.4 Hz, 1F), -103.4 (dd, JFF = 249.1 Hz, JHF = 13.0 Hz, 1F). EMAR (EI +) calculated for C25H25F2IN2O3 [M +]: 566.0878, found: 566.0865.

(2R, 3R) -1-Phenyl-1,1-di-fluoro-3-hydroxy-4- (3-iodobenzylamino) butaneyl-2-carbamate benzyl (17b). Following the same procedure above, 112 mg of epoxy 12b provided 142 mg of 17b (75%). Yellow solid, mp: 109-111 ° C (CH2Cl2). [α] D25 = -16.8 (c 1.0, CHCl3). 1H-NMR (300 MHz, CDCl3) δ 2.69 (dd, J = 12.6, 5.4 Hz, 1H),

2.75 (dd, J = 12.6, 3.9 Hz, 1H), 3.53 (d, J = 13.5 Hz, 1H), 3.59 (d, J = 13.5 Hz, 1H), 3.81 (dd, J = 5.4, 3.9 Hz, 1H ), 4.34-4.48 (m, 1H), 4.86 (d, J = 12.3 Hz, 1H), 4.97 (d, J = 12.3 Hz, 1H), 6.25 (d, J = 10.3 Hz, 1H), 6.90 (t , J = 7.8 Hz, 1H), 7.10-7.54 (m, 13H). 13C-NMR (75.5 MHz, CDCl3) δ 49.8, 52.1, 58.9 (t, JCF = 28.0 Hz), 66.1, 66.4, 93.5, 120.4 (t, JCF = 248.4 Hz), 124.4 (t, JCF = 6.4 Hz), 126.3, 126.9, 127.1, 127.4, 127.5, 129.2, 129.2, 133.8 (t, JCF = 25.7 Hz), 135.1, 135.2, 136.0, 140.9, 155.7. 19F-NMR (CDCl3, 282.4 MHz) δ -104.9 (dd, JFF = 250.2 Hz, JHF = 11.6 Hz, 1F), -102.0 (dd, JFF = 250.2 Hz, JHF = 15.8 Hz, 1F). EMAR (EI +) calculated for C25H25F2IN2O3 [M +]: 566.0878, found: 566.0876.

(2R, 3R) -1-Phenyl-1,1-di-fluoro-3-hydroxy-4- (3-iodobenzylamino) benzyl butanyl-2-carbamate (17'a) .50mg of 12'a epoxide provided 58 mg (68 %) of (17'a) after fl ash chromatography. [α] D25 = +18.80 (c 1.0, CHCl3).

(2S, 3S) -1-Phenyl-1,1-di-fluoro-3-hydroxy-4- (3-iodobenzylamino) benzyl butanyl-2-carbamate (17'b) .36mg of 12'b epoxide led, after chromatography , at 43 mg (70%) of (17'b), [α] D25 = +15.60 (c 1.0, CHCl3).

(2R, 3S) -1-Phenyl-1,1-di-fluoro-3-hydroxy-4- (3-iodobenzylamino) butanyl-2-carbamate tert-butyl ester (16a). To a solution of epoxide 13a (48 mg, 0.16 mmol) in anhydrous isopropanol (1.0 mL) at room temperature 3-iodobenzylamine (0.18 mmol) was added and the mixture was heated at 70 ° C with constant stirring, until it was observed the disappearance of the starting epoxide by CCF. Then, the solvent was removed under reduced pressure and the crude obtained was

purified by fl ash chromatography, using n-hexane: AcOEt (2: 1) as eluent, 74 mg of amino alcohol 16a (87%) being isolated as a clear oil. [α] D25 = -17.99 (c 1.0, CHCl3). 1H-NMR (300 MHz, CDCl3) δ 1.34 (s, 9H),

2.58 (dd, J = 12.0, 9.0 Hz, 1H), 2.67 (dd, J = 12.0, 4.5 Hz, 1H), 3.65 (d, J = 13.5 Hz, 1H), 3.72 (d, J = 13.5 Hz, 1H ), 4.01 (dd, J = 9.0, 4.5 Hz, 1H), 4.06-4.16 (m, 1H), 5.28 (d, J = 10.2 Hz, 1H), 7.03 (t, J = 7.5 Hz, 1H), 7.19 (d, J = 7.5 Hz, 1H), 7.41-7.62 (m, 7H). 13C-NMR (75.5 MHz, CDCl3) δ 28.1, 51.4, 52.6, 56.5 (t, JCF = 28.2 Hz), 65.7, 79.9, 94.4, 121.8 (t, JCF = 249.3 Hz), 125.6 (t, JCF = 6.3 Hz ), 127.2, 128.2, 130.0, 130.2, 134.8 (t, JCF = 25.8 Hz), 136.2, 136.9, 142.0,

155.5. 19F-NMR (CDCl3, 282.4 MHz) δ -104.3 (dd, JHF = 14.4 Hz, 1F), -104.3 (dd, JHF = 13.0 Hz, 1F). EMAR (FAB) calculated for C22H28F2IN2O3 [M + H +]: 533.1113, found: 533.1092.

(2R, 3R) -1-Phenyl-1,1-di-fluoro-3-hydroxy-4- (3-iodobenzylamino) butanyl-2-carbamate tert-butyl (16b). 21.5 mg of epoxide 13b (0.07 mmol) led to 16b (35.6 mg, 93%). White solid, mp: 100-102 ° C (CH2Cl2). [α] D25 = -10.25 (c 1.0, CHCl3). 1H-NMR (300 MHz, CDCl3) δ 1.22 (sa, 1H), 1.34 (s, 9H), 2.31 (sa, 1H), 2.75-2.84 (m, 2H),

3.69 (d, J = 13.8 Hz, 1H), 3.74 (d, J = 13.8 Hz, 1H), 3.89 (dd, J = 9.2, 4.7 Hz, 1H), 4.32-4.49 (m, 1H), 5.46 (d , J = 9.6 Hz, 1H), 7.05 (t, J = 7.5 Hz, 1H), 7.26 (d, J = 7.5 Hz, 1H), 7.38-7.45 (m, 3H), 7.45-7.53 (m, 2H) , 7.58 (d, J = 7.9 Hz, 1H), 7.65 (s, 1H). 13C-NMR (75.5 MHz, CDCl3) δ 28.1, 50.8, 53.1, 58.9 (t, JCF = 27.2 Hz), 67.9, 80.1, 94.4, 121.6 (t, JCF = 248.9 Hz), 125.4 (t, JCF = 6.0 Hz ), 127.3, 128.3, 130.0, 130.2, 135.0 (t, JCF = 25.4 Hz), 136.1, 137.0, 142.2, 155.8. 19F-NMR (CDCl3, 282.4 MHz) δ -104.6 (dd, JFF = 249.6 Hz, JHF = 11.6 Hz, 1F), -101.8 (dd, JFF = 249.6 Hz, JHF = 16.4 Hz, 1F). EMAR (FAB) calculated for C22H28F2IN2O3 [M + H +]: 533.1113, found: 533.1113.

(2S, 3R) -1-Phenyl-1,1-di-fluoro-3-hydroxy-4- (3-iodobenzylamino) butanyl-2-carbamate tert-butyl (16'a) .35mg of epoxide 13'a led to 53 mg, 89% of 16'a [α] D25 = +22.57 (c 1.0, CHCl3).

(2S, 3S) -1-Phenyl-1,1-di-fluoro-3-hydroxy-4- (3-iodobenzylamino) butanyl-2-carbamate tert-butyl (16'b) .67mg of epoxide 13'b led to 74 mg, 62% of 16'b. [α] D25 = +11.32 (c 1.0, CHCl3).

N- (4-Bromophenyl) sulfonyl [(1R) -1- (phenyldi fl uoromethyl) allyl] amine (9). To a solution of allylamine 4 (100 mg, 0.3 mmol) in anhydrous methanol (2.2 mL) was added 0.87 mL of a solution of HCl (4 M) in dioxane, and allowed to stir at room temperature for 2 hours After this time, the reaction crude was concentrated to dryness and the white residue obtained was redissolved in 2.2 mL of dioxane. Then, K2CO3 (1.0 mmol), (4-bromophenyl) sulfonyl chloride (1.7 mmol) and a catalytic amount of dimethylaminopyridine (DMAP) (0.05 mmol) were successively added at room temperature. The reaction mixture was left with continuous stirring at room temperature for 18 hours. After this period, the resulting mixture was hydrolyzed with brine, the two phases were separated and the aqueous phase was extracted with AcOEt. The combined organic phases were washed with brine and dried over Na2SO4. After evaporating the solvent under reduced pressure, the oil obtained was subjected to fl ash chromatography, using n-hexane: ethyl ether (6: 1) as eluent, obtaining a white solid corresponding to allylamine 9 (92 mg, 66%) as a white solid, mp: 94-96 ° C (CH2Cl2). [α] D25 = +4.12 (c 1.0, CHCl3). 1H-NMR (300 MHz, CDCl3) δ 4.47 (tddt, J = 11.6, 9.6, 6.0, 2.1 Hz, 1H), 5.13 (d, J = 9.6 Hz, 2H), 5.24 (d, J = 18.0 Hz, 1H ), 5.25 (d, J = 10.2 Hz, 1H), 6.75 (ddd, J = 18.0, 10.2, 6.0 Hz, 1H), 7.40-7.43 (m, 3H), 7.47-7.50 (m, 2H). 13C-NMR (75.5 MHz, CDCl3) δ 61.4 (t, JCF = 30.6 Hz), 120.2 (t, JCF = 249.4 Hz), 120.6, 125.7 (JCF = 6.2 Hz), 127.5, 128.4, 128.4, 130.1 (t, JCF = 2.9 Hz), 130.3, 132.1, 133.5 (t, JCF = 25.7 Hz), 139.6. 19F-NMR (CDCl3, 282.4 MHz) δ -104.4 (d, JHF = 11.6 Hz, 2F). EMAR (EI +) calculated for C16H14BrF2NO2S [M +]: 400.9897, found: 400.9904.

(2R, 3S) -N- {2- [1-Phenyl-1,1-di-fluoro-4- (3- fl uorophenylmethyl) amino-3-hydroxy]} benzyl carbamate (18a).

Isomer (2R, 3S). To a solution of the epoxide (49.8 mg, 0.15 mmol) in anhydrous isopropanol (2.0 mL) was added, at room temperature, the primary amine (3-fl uorophenylmethyl) amine (0.34 mmol) and molecular sieve (3

TO). The reaction mixture was heated in a sealed tube for 16 hours at 70 ° C with constant stirring until the starting compound (TLC) disappeared. Then, the solvent was removed under reduced pressure and the crude was purified by fl ash chromatography using hexane: ethyl acetate (2: 1) and 1% methanol to obtain product 18a (51.8 mg) as a white solid with a white solid. 76% yield Mp: 77-81 ° C. [α] D25 = -23.68 (c 1.0, CHCl3). 1H-NMR (CDCl3, 300 MHz) δ 1.71 (sa, 2H), 2.53-2.60 (m, 2H), 2.65-2.70 (m, 2H), 3.71 (s, 2H), 3.90 (dd J = 9.6, 4.2 Hz, 1H), 4.10

4.23 (m, 1H), 5.00 (d, J = 12.4 Hz, 2H), 5.55 (d, J = 9.9 Hz, 1H), 6.91-7.01 (m, 3H), 7.22-7.28 (m, 3H), 7.27 -7.28 (m, 1H), 7.31-7.35 (m, 2H), 7.40-7.44 (m, 3H), 7.49-7.51 (m, 2H). 13C-NMR (75.5 MHz, CDCl3) δ 51.3, 52.7, 57.1 (t, JCF = 28.1 Hz), 65.6, 67.0, 114.0 (d, JCF = 21.4 Hz), 114.7 (d, JCF = 21.2 Hz), 121.6 ( t, JCF = 248.9 Hz), 123.4 (d, JCF =

2.7 Hz), 125.5 (t, JCF = 6.3 Hz), 127.8, 128.1, 128.4 (d, JCF = 8.9 Hz), 129.9 (d, JCF = 8.2 Hz), 130.7, 134.5 (t, JCF = 25.7 Hz), 136.1, 142.0 (d, JCF = 6.7 Hz), 156.3, 161.2, 164.5. 19F-NMR (CDCl3, 282.4 MHz) δ -103.4 (dd, JFF = 249.2 Hz, JHF = 14.2 Hz, 1F), -104.6 (d, JFF = 248.9 Hz, JHF = 13.2 Hz, 1F), -113.6 (tdd , JCF = 24.2, 9.0, 6.0 Hz, 1F). EMAR (ESI +) calculated for C25H25F3N2O3 [M + H +]: 459.1896, found: 459.1899.

(2R, 3R) -N- {2- [1-Phenyl-1,1-di-fluoro-4- (3- fl uorophenylmethyl) amino-3-hydroxy]} benzyl carbamate (18b).

Isomer (2R, 3R). To a solution of the epoxide (47.5 mg, 0.14 mmol) in anhydrous isopropanol (1.9 mL) was added, at room temperature, the primary amine (3-fl uorophenylmethyl) amine (0.32 mmol) and molecular sieve (3

TO). The reaction mixture was heated in a sealed tube for 24 hours at 70 ° C with constant stirring until the starting compound (TLC) disappeared. Then, the solvent was removed under reduced pressure and the crude was purified by fl ash chromatography using hexane: ethyl acetate (2: 1) and 1% methanol to obtain product 18b (38.7 mg) as a white solid with a white solid. 61% yield. Mp: 134-137 ° C. [α] D25 = -20.51 (c 1.0, CHCl3). 1H-NMR (CDCl3, 300 MHz) δ 2.1 (sa, 2H), 2.76-2.88 (m, 2H), 3.74 (d, J = 13.8 Hz, 2H), 3.90 (dd J = 9.9, 5.7 Hz, 1H) , 4.43-4.57 (m, 1H), 4.27-4.58 (m, 1H), 5.02 (d, J = 12.3 Hz, 2H), 5.06 (s, 2H), 7.26-7.56 (m, 10H). 13C-NMR (CDCl3, 75.5 MHz) δ 51.0, 53.7, 60.0 (t, JCF = 26.5 Hz), 67.5, 68.1, 114.4 (d, JCF = 21.2 Hz), 115.2 (d, JCF = 21.1 Hz), 121.9 ( t, JCF = 251.6 Hz),

123.9 (d, JCF = 2.8 Hz), 125.8 (t, JCF = 6.3 Hz), 128.3, 128.5, 128.8, 128.9, 130.3 (d, JCF = 8.3 Hz), 130.6, 134.2, 135.1 (t, JCF = 26.3 Hz ), 136.4, 142.6, 156.9. 19F-NMR (CDCl3, 282.4 MHz) δ -101.89 (dd, JFF = 251.0 Hz, JHF = 15.8 Hz, 1F), -104.8 (d, JFF = 250.2 Hz, JHF = 12.1 Hz, 1F), -113.7 (tdd , JCF = 28.4, 8.9, 6.1, 2.8 Hz, 1F). EMAR (FAB) calculated for C25H25F3N2O3 [M + H +]: 458.1817, found: 458.1810.

(2R, 3S) -N- {2- [4- (3-Chlorophenylmethyl) amino-1-phenyl-1,1-di-fluoro-3-hydroxy]} benzyl carbamate (19a).

Isomer (2R, 3S). To a solution of the epoxide (43.1 mg, 0.13 mmol) in anhydrous isopropanol (1.8 mL) was added, at room temperature, the primary amine (3-chlorophenylmethyl) amine (0.19 mmol) and molecular sieve (3

TO). The reaction mixture was heated in a sealed tube for 8 hours at 70 ° C with constant stirring until the starting compound (TLC) disappeared. Then, the solvent was removed under reduced pressure and the crude was purified by fl ash chromatography using hexane: ethyl acetate (1: 1) to obtain product 19a (45.6 mg) as a white solid with a 74% yield. Mp: 89-92 ° C. [α] D25 = -24.89 (c 1.0, CHCl3). 1H-NMR (300 MHz, CDCl3) δ 2.32 (sa, 2H), 2.56-2.72 (m, 2H), 3.71 (s, 2H), 4.02 (dd, J = 4.2 Hz, 1H), 4.14-4.28 (m , 1H), 5.07 (d, J = 12.3 Hz, 2H), 5.61 (d, J = 9.9 Hz, 1H), 7.11-7.15 (m, 1H), 7.24-7.31 (m, 5H), 7.34-7.38 ( m, 3 H), 7.40-7.49 (m, 3 H), 7.52, 7.55 (m, 2H). 13C-NMR (75.5 MHz, CDCl3) δ 51.4, 52.7, 57.2 (t, JCF = 28.5 Hz), 65.6, 67.0, 121.6 (t, JCF = 248.5 Hz), 125.5 (t, JCF =

6.4 Hz), 126.0, 127.3, 127.8, 128.0, 128.1, 128.3, 128.4, 129.7, 130.1, 134.3, 134.6 (t, JCF = 25.7 Hz), 141.6, 156.3. 19F-NMR (CDCl3, 282.4 MHz) δ -103.4 (dd, JFF = 249.0 Hz, JHF = 14.6 Hz, 1F), -104.6 (dd, JFF = 249.0 Hz, JHF = 13.2 Hz, 1F). EMAR (EI +) calculated for C25H25ClF2N2O3 [M +]: 474.1522, found: 474.1518.

(2R, 3R) -N- {2- [4- (3-Chlorophenylmethyl) amino-1-phenyl-1,1-di-fluoro-3-hydroxy]} benzyl carbamate (19b).

Isomer (2R, 3R). To a solution of the epoxide (46.4 mg, 0.14 mmol) in anhydrous isopropanol (1.9 mL) was added, at room temperature, the primary amine (3-chlorophenylmethyl) amine (0.31 mmol) and molecular sieve (3

TO). The reaction mixture was heated in a sealed tube for 24 hours at 70 ° C with constant stirring until the starting compound (TLC) disappeared. Then, the solvent was removed under reduced pressure and the crude was purified by fl ash chromatography using hexane: ethyl acetate (2: 1) and 1% methanol to obtain product 19b (39.9 mg) as a white solid with a white solid. 60% yield Mp: 120-123 ° C. [α] D25 = -17.3 (c 1.0, CHCl3). 1H-NMR (300 MHz, CDCl3) δ 1.60 (sa, 2H), 2.76-2.87 (m, 2H), 3.71 (d, J = 13.5 Hz, 1H), 3.81 (dd, J = 9.4, 5.4 Hz, 1H ), 4.43-4.57 (m, 1H), 5.06 (d, J = 12.3 Hz, 2H), 5.99 (d, J = 9.9 Hz, 1H), 6.91-7.05 (m, 3H), 7.21-7.50 (m, 11H). 13C-NMR (75.5 MHz, CDCl3) δ 51.0, 53.6, 60.1 (t, JCF = 28.7 Hz), 67.5, 68.0, 121.8 (t, JCF = 251.4 Hz), 125.8 (t, JCF = 6.4 Hz), 126.5, 127.7, 128.3, 128.5, 128.8, 128.9, 130.1, 130.6, 134.6, 135.7 (t, JCF = 25.4 Hz), 136.5, 142.1, 157.0. 19F-NMR (CDCl3, 282.4 MHz) δ -101.8 (dd, JFF = 250.3 Hz, JHF = 16.0 Hz, 1F), -104.8 (dd, JFF = 250.4 Hz, JHF = 11.2 Hz, 1F), -113.7. EMAR (EI +) calculated for C25H25ClF2N2O3 [M +]: 474.1522, found: 474.1516.

(2R, 3R) -N- {2- [1-Phenyl-1,1-di-fluoro-3-hydroxy-4- (3-methoxyphenylmethyl) amino]} benzyl carbamate (20b).

Isomer (2R, 3R). To a solution of the epoxide (42.4 mg, 0.34 mmol) in anhydrous isopropanol (1.7 mL) was added, at room temperature, the primary amine (3-methoxyphenylmethyl) amine (0.48 mmol) and molecular sieve (3

TO). The reaction mixture was heated in a sealed tube for 24 hours at 70 ° C with constant stirring until the starting compound (TLC) disappeared. Then, the solvent was removed under reduced pressure and the crude was purified by fl ash chromatography using hexane: ethyl acetate (1: 2) and 1% to obtain product 20b (50.0 mg) as a 84% white solid. of performance. Mp: 101-105 ° C. [α] D25 = -18.95 (c 1.0, CHCl3). 1H-NMR (CDCl3, 300 MHz) δ 1.59 (sa, 2H), 2.76-2.90 (m, 2H), 3.71 (d, J = 15.0 Hz, 2H), 3.77 (s, 3H), 3.86-3.91 (m , 1H), 4.41-4.57 (m, 1H), 5.03 (d J = 12.3 Hz, 2H), 5.86 (d, J = 10.2 Hz, 1H), 6.78-6.90 (m, 3H), 7.18-7.24 (m , 2H), 7.32-7.50 (m, 9H). 13C-NMR (CDCl3, 75.5 MHz) δ 50.6, 53.7, 55.1 59.7, (t, JCF = 28.5 Hz), 67.0, 67.4, 112.6, 113.4, 121.7 (t, JCF = 248.3 Hz), 120.3,

125.4 (t, JCF = 6.3 Hz), 127.8, 128.0, 128.3, 128.4, 129.4, 130.1, 134.8 (t, JCF = 25.5 Hz), 136.1, 141.1, 156.5, 159.7. 19F-NMR (CDCl3, 282.4 MHz) δ -1041.4 (dd, JFF = 250.7 Hz, JHF = 14.6 Hz, 1F), -104.9 (dd, JFF = 250.7 Hz, JHF = 12.1 Hz, 1F). EMAR (ESI +) calculated for C26H28F2N2O4 [M + H +]: 471.2095, found: 471.2090.

(2R, 3R) -N- {2- [1-Phenyl-1,1-di-fluoro-3-hydroxy-4- (2-thienylmethyl) amino]} benzyl carbamate (21b).

Isomer (2R, 3R). To a solution of the epoxide (49.0 mg, 0.12 mmol) in anhydrous isopropanol (2.0 mL) was added, at room temperature, the primary amine (2-thienylmethyl) amine (0.26 mmol) and molecular sieve (3

TO). The reaction mixture was heated in a sealed tube for 21 hours at 70 ° C with constant stirring until the starting compound (TLC) disappeared. Then, the solvent was removed under reduced pressure and the crude was purified by fl ash chromatography using hexane: ethyl acetate (2: 1) to obtain product 21b (43.3 mg) as a white solid in 83% yield. Mp: 133-136 ° C. [α] D25 = -20.10 (c 1.0, CHCl3). 1H-NMR (300 MHz, CDCl3) δ

1.60 (sa, 2H), 2.76-2.87 (m, 2H), 3.71 (d, J = 13.5 Hz, 1H), 3.81 (dd, J = 9.4, 5.4 Hz, 1H), 4.43-4.57 (m, 1H) , 5.06 (d, J = 12.3 Hz, 2H), 5.99 (d, J = 9.9 Hz, 1H), 6.91-7.05 (m, 3H), 7.21-7.50 (m, 11H). 13C-NMR (75.5 MHz, CDCl3) δ 48.2, 50.4, 59.6 (t, JCF = 27.4 Hz), 67.0, 67.6, 121.3, (t, JCF = 248.4 Hz), 124.5, 125.0, 125.4 (t, JCF = 6.3 Hz), 126.6, 127.8, 128.0, 128.3, 128.4, 130.1, 134.8 (t, JCF = 25.6 Hz), 136.1, 143.2, 156.5. 19F-NMR (CDCl3, 282.4 MHz) δ -101.9 (dd, JFF = 250.4 Hz, JHF = 16.0 Hz, 1F), -104.7 (dd, JFF = 250.3 Hz, JHF = 11.8 Hz, 1F). EMAR (ESI +) calculated for C23H24F2N2O3S [M + H +]: 447.1554, found: 447.1552.

(2R, 3R) -N- {2- [1-Phenyl-1,1-di-fluoro-4- (2-furylmethyl) amino-3-hydroxy]} benzyl carbamate (22b).

Isomer (2R, 3R). To a solution of the epoxide (48.7 mg, 0.15 mmol) in anhydrous isopropanol (2.0 mL) was added, at room temperature, the primary amine (2-furylmethyl) amine (0.33 mmol) and molecular sieve (3

TO). The reaction mixture was heated in a sealed tube for 24 hours at 70 ° C with constant stirring until the starting compound (TLC) disappeared. Then, the solvent was removed under reduced pressure and the crude was purified by fl ash chromatography using pentane: ethyl ether (4: 6) to obtain product 22b (40.7 mg) as a white solid with a 65% yield. Mp: 135-140 ° C. [α] D25 = -19.16 (c 1.0, CHCl3). 1H-NMR (300 MHz, CD2Cl2) δ 2.26 (sa, 2H), 2.69-2.76 (m, 1H), 2.80-2.85 (m, 1 H), 3.79-3.84 (m, 1H), 4.37-4.52 (m , 1H), 4.99 (d, J = 12.4 Hz, 1H), 5.80 (d, J = 9.6 Hz, 1H), 6.23 (d, 3.3 Hz, 1H), 6.31-6.32 (m, 1H), 7.22-7.26 (m, 2H), 7.29-7.33 (m, 4H), 7.39-7.49 (m, 5H) .13C-NMR (75.5 MHz, CDCl3) δ 46.6, 51.1, 54.4 (t, JCF = 27.4 Hz), 67.6, 68.2, 107.8, 110.9, 122.2 (t, JCF = 253.0 Hz),

126.2 (t, JCF = 6.4 Hz), 128.5, 128.8, 129.1, 129.2, 130.9, 135.7 (t, JCF = 25.6 Hz), 137.3, 142.6, 154.4, 157.1. 19F-NMR (CDCl3, 282.4 MHz) δ -101.3 (dd, JFF = 249.9 Hz, JHF = 16.0 Hz, 1F), -104.2 (dd, JFF = 249.6 Hz, JHF = 11.8 Hz, 1F). EMAR (ESI +) calculated for C23H24F2N2O4 [M + H +]: 431.1782, found: 431.1781.

(2R, 3S) -N- {2- [1-Phenyl-1,1-di-fluoro-4- (3- fl uorophenylmethyl) amino-3-hydroxy]} tert-butyl carbamate (23a).

Isomer (2R, 3S). To a solution of the epoxide (50 mg, 0.17 mmol) in anhydrous isopropanol (2.1 mL) was added, at room temperature, the primary amine (3-fl uorophenylmethyl) amine (0.38 mmol) and molecular sieve (3 ˚

TO). The reaction mixture was heated in a sealed tube for 6 hours at 70 ° C with constant stirring until the starting compound (TLC) disappeared. Then, the solvent was removed under reduced pressure and the crude was purified by fl ash chromatography using hexane: ethyl acetate (2: 1) to obtain product 23a (53.2 mg) as a yellow oil in 75% yield. [α] D25 = -21.9 (c 1.0, CHCl3). 1H-NMR (300 MHz, CDCl3) δ 1.33 (s, 9H), 2.55 (sa, 2H), 2.64-2.69 (m, 2H), 3.73 (d, J = 13.5 Hz, 2H), 4.01-4.05 (m , 1H), 4.06-4.16 (m, 1H), 5.33 (d, J = 9.9 Hz, 1H), 6.90-7.02 (m, 3H), 7.21-7.29 (m, 1H), 7.38-7.42 (m, 3H ), 7.47-7.52 (m, 2H). 13C-NMR (75.5 MHz, CDCl3) δ 28.5, 51.8, 53.2, 56.9 (t, JCF = 27.9 Hz), 66.1, 80.3, 114.4 (d, JCF = 21.1 Hz), 115.2 (d, JCF = 21.2 Hz), 122.4 (t, JCF = 248.8 Hz), 123.9 (d, JCF = 2.7 Hz), 126.0 (t, JCF = 6.3 Hz), 128.6, 130.34 (d, JCF = 8.4 Hz), 135.2 (t, JCF = 25.7 Hz ), 142.4 (d, JCF = 6.8 Hz), 155.9, 161.6, 164.9. 19F-NMR (CDCl3, 282.4 MHz) δ -104.3 (dd, JHF = 19.7, 5.6 Hz, 2 F), -113.6 (td, JCF = 24.5, 9.2, 6.0 Hz, 1 F). EMAR (ESI +) calculated for C22H27F3N2O3 [M + H +]: 425.2052, found: 425.2053.

(2R, 3R) -N- {2- [1-Phenyl-1,1-di-fluoro-4- (3- fl uorophenylmethyl) amino-3-hydroxy]} tert-butyl carbamate (23b).

Isomer (2R, 3R). To a solution of the epoxide (53.2 mg, 0.18 mmol) in anhydrous isopropanol (2.2 mL) was added, at room temperature, the primary amine (3-fl uorophenylmethyl) amine (0.67 mmol) and molecular sieve (3

TO). The reaction mixture was heated in a sealed tube for 24 hours at 70 ° C with constant stirring until the starting compound (TLC) disappeared. Then, the solvent was removed under reduced pressure and the crude was purified by fl ash chromatography using hexane: ethyl acetate (2: 1) to obtain product 23b (48.9 mg) as a white solid in 65% yield. Mp: 101-104 ° C. [α] D25 = -12.34 (c 1.0, CHCl3). 1H-NMR (300 MHz, CDCl3) δ 1.33 (s, 9H), 2.48 (sa, 2H), 2.82 (m, 2H), 3.76 (d, J = 13.5 Hz, 2H), 3.89 (dd, J = 9.6 , 5.1 Hz, 1H), 4.32-4.47 (m, 1H), 5.59 (d, J = 9.9 Hz, 1H), 6.91-7.07 (m, 4H), 7.23-7.28 (m, 1H), 7.40-7.50 ( m, 4H). 13C-NMR (75.5 MHz, CDCl3) δ 28.1, 50.8, 53.3, 58.9 (t, JCF = 26.9 Hz), 67.9, 80.1, 113.0 (d, JCF = 21.2 Hz), 114.8 (d, JCF = 21.2 Hz), 121.6 (t, JCF = 251.5 Hz), 123.5 (d, JCF = 2.7 Hz), 125.4 (t, JCF = 6.3 Hz), 128.3, 130.3 (d, JCF = 8.1 Hz), 135.0 (t, JCF = 25.7 Hz ), 142.4 (d, JCF = 6.7 Hz), 155.7, 161.3, 164.5. 19F-NMR (CDCl3, 282.4 MHz) δ -102.7 (dd, JHF = 249.5, 16.3 Hz, 1F), -104.2 (dd, JHF = 249.3, 11.5 Hz, 1F), -113.7 (td, JCF = 28.2, 8.9 , 6.1 Hz, 1F). EMAR (FAB) calculated for C22H27F3N2O3 [M + H +]: 425.2052, found: 425.2047.

(2R, 3S) -N- {2- [1-Phenyl-1,1-di-fluoro-3-hydroxy-4- (2-thienylmethyl) amino]} tert-butyl carbamate (24a).

Isomer (2R, 3S). To a solution of the epoxide (50.0 mg, 0.78 mmol) in anhydrous isopropanol (2.1 mL) was added, at room temperature, the primary amine (2-thienylmethyl) amine (0.38 mmol) and molecular sieve (3

TO). The reaction mixture was heated in a sealed tube for 24 hours at 70 ° C with constant stirring until the starting compound (TLC) disappeared. Then, the solvent was removed under reduced pressure and the crude was purified by fl ash chromatography using hexane: ethyl acetate (2: 1) and 1% methanol to obtain product 24a (45.0 mg) as a yellow oil with a 65% yield. [α] D25 = -20.90 (c 1.0, CHCl3). 1H-NMR (300 MHz, CDCl3) δ

1.34 (s, 9H), 2.52 (sa, 2H), 2.68-2.74 (m, 2), 3.95 (d, J = 14.4 Hz, 2H), 3.99-4.15 (m, 2H), 5.33 (d, J = 9.9 Hz, 1H), 6.86-6.93 (m, 2H), 7.19 (dd, J = 4.9, 1.2 Hz, 1H), 7.40-7.52 (m, 5H). 13C-NMR (75.5 MHz, CDCl3) δ 28.1, 47.7, 51.1,

56.5 (t, JCF = 27.9 Hz), 65.6, 79.8, 121.6 (t, JCF = 251.9 Hz), 124.6, 125.1, 125.6 (t, JCF = 6.4 Hz), 126.6, 128.2, 129.9,

134.8 (t, JCF = 25.7 Hz), 143.0, 155.4. 19F-NMR (CDCl3, 282.4 MHz) δ -104.3 (d, JHF = 14.2 Hz, 2F). EMAR (FAB) calculated for C20H26F2N2O3S [M + H +]: 413.1710, found: 413.1715.

(2R, 3R) -N- {2- [1-Phenyl-1,1-di-fluoro-3-hydroxy-4- (2-thienylmethyl) amino]} tert-butyl carbamate (24b).

Isomer (2R, 3R). To a solution of the epoxide (50.8 mg, 0.17 mmol) in anhydrous isopropanol (2.1 mL) was added, at room temperature, the primary amine (2-thienylmethyl) amine (0.51 mmol) and molecular sieve (3

TO). The reaction mixture was heated in a sealed tube for 24 hours at 70 ° C with constant stirring until the starting compound (TLC) disappeared. Then, the solvent was removed under reduced pressure and the crude was purified by fl ash chromatography using hexane: ethyl acetate (2: 1) and 1% methanol to obtain product 24b (48.5 mg) as a 69% white solid. of performance. Mp: 95-97 ° C. [α] D25 = -17.26 (c 1.0, CHCl3). 1H-NMR (300 MHz, CDCl3) δ 1.21 (sa, 2 H), 1.33 (s, 9H), 2.79-2.87 (m, 2H), 3.85-3.90 (m, 1H), 3.97, (d, J = 14.4 Hz, 2H), 4.32-4.47 (m, 1H), 5.37 (d, J = 9.9 Hz, 1H), 6.90-6.95 (m, 2H), 7.21 (dd, J = 6.3, 1.2 Hz, 1H), 7.40-7.42 (m, 4 H), 7.48-7.5 (m, 2 H). 13C-NMR (75.5 MHz, CDCl3) δ 28.1, 48.2, 50.5, 55.7, 58.7 (t, JCF = 28.4 Hz), 68.0, 80.1, 121.6 (t, JCF = 251.1 Hz), 124.4, 124.9, 125.4 (t, JCF = 6.4 Hz), 126.6, 128.3, 130.0, 135.0 (t, JCF = 25.4 Hz), 143.4, 155.6. 19F-NMR (CDCl3,

282.4 MHz) δ -102.2 (dd, JHF = 247.9, 16.3 Hz, 1F), -104.5 (dd, JHF = 249.9, 11.8 Hz, 1F). EMAR (FAB) calculated for C20H26F2N2O3S [M + H +]: 412.1632, found: 412.1623.

(2R, 3R) -N- {2- [4- (3-Chlorophenylmethyl) amino-1-phenyl-1,1-di-fluoro-3-hydroxy]} tert-butyl carbamate (25b).

Isomer (2R, 3R). To a solution of the epoxide (50.1 mg, 0.17 mmol) in anhydrous isopropanol (2.1 mL) was added, at room temperature, the primary amine (3-chlorophenylmethyl) amine (0.63 mmol) and molecular sieve (3

TO). The reaction mixture was heated in a sealed tube for 24 hours at 70 ° C with constant stirring until the starting compound (TLC) disappeared. Then, the solvent was removed under reduced pressure and the crude was purified by fl ash chromatography using hexane: ethyl acetate (2: 1) to obtain product 25b (42.9 mg) as a white solid in 58% yield. Mp: 110-113 ° C. [α] D25 = -12.34 (c 1.0, CHCl3). 1H-NMR (300 MHz, CDCl3) δ

1.33 (s, 9H), 2.40 (sa, 2H), 2.81 (m, 2H), 3.74 (d, J = 13.6 Hz, 2H), 3.89 (d, J = 9.9, 5.2 Hz, 1H), 4.32-4.46 (m, 1H),

5.51 (d, J = 9.6 Hz, 1H), 7.16-7.19 (m, 1H), 7.23-7.25 (m, 2H), 7.40-7.42 (m, 3H), 7.47-7.50 (m, 2H). 13C-NMR

(75.5 MHz, CDCl3) δ 28.1, 50.8, 53.3, 58.9 (t, JCF = 26.5 Hz), 68.0, 80.1, 121.6 (t, JCF = 248.5 Hz), 125.4 (t, JCF = 6.2 Hz), 126.1, 127.2 , 128.1, 128.3, 129.6, 130.0, 134.2, 135.0 (t, JCF = 25.4 Hz), 141.8, 155.7. 19F-NMR (CDCl3, 282.4 MHz) δ -104.8 (dd, JHF = 250.2, 11.5 Hz, 1F), -105.5 (dd, JHF = 250.2, 14.6 Hz, 1F). EMAR (FAB) calculated for C22H27FClN2O3 [M + H +]: 440.1678, found: 439.0976.

(2R, 3R) -N- {2- [1-Phenyl-1,1-di-fluoro-3-hydroxy-4- (3-methoxyphenylmethyl) amino]} tert-butyl carbamate (26b).

Isomer (2R, 3R). To a solution of the epoxide (54.9 mg, 0.18 mmol) in anhydrous isopropanol (2.3 mL) was added, at room temperature, the primary amine (3-methoxyphenylmethyl) amine (0.55 mmol) and molecular sieve (3

TO). The reaction mixture was heated in a sealed tube for 24 hours at 70 ° C with constant stirring until the starting compound (TLC) disappeared. Then, the solvent was removed under reduced pressure and the crude was purified by fl ash chromatography using hexane: ethyl acetate (2: 1) and 1% methanol to obtain product 26b (17.1 mg) as a yellow oil with 21% of performance. [α] D25 = -12.34 (c 1.0, CHCl3). 1H-NMR (300 MHz, CDCl3) δ

1.33 (s, 9H), 2.21 (sa, 2H), 2.82 (m, 2H), 3.74 (d, J = 13.5 Hz, 2H), 3.83 (s, 3H), 3.88 (dd, J = 9.6, 5.4 Hz , 1H), 4.32

4.47 (m, 1H), 5.39 (d, J = 9.9 Hz, 1H), 6.78-6.89 (m, 3H), 7.20-7.26 (m, 1 H), 7.40-7.51 (m, 5 H). 13C-NMR (75.5 MHz, CDCl3) δ 28.1, 50.7, 53.7, 55.1, 58.7 (t, JCF = 26.7 Hz), 67.9, 80.0, 112.4, 113.5, 121.5 (t, JCF = 249.9 Hz), 120.2,

125.4 (t, JCF = 6.3 Hz), 128.2, 129.4, 130.0, 135.0 (t, JCF = 26.1 Hz), 141.3, 155.6, 159.7. 19F-NMR (CDCl3, 282.4 MHz) δ -102.3 (dd, JHF = 254.1, 16.0 Hz, 1F), -104.4 (dd, JHF = 254.1, 11.2 Hz, 1F). EMAR (FAB) calculated for C23H30F2N2O4 [M + H +]: 436.2174, found: 436.2127.

(2R, 3S) -1-Phenyl-1,1-di-fluoro-3-hydroxy-4- (dipropylamino) butane-2-carbamate benzyl (27a). To a solution of epoxide 12a (68 mg, 0.20 mmol) in anhydrous isopropanol (0.7 mL) at room temperature dipropylamine (0.3 mmol) was added and the mixture was heated at 70 ° C with constant stirring, until it was observed the disappearance of the starting epoxide by CCF. Then, the solvent was removed under reduced pressure and the crude obtained was purified by fl ash chromatography, using n-hexane: AcOEt (2: 1) as eluent, the product 27a (72 mg, 81%) being isolated as a yellowish solid. Mp = 69-71 ° C (CH2Cl2). [α] D25 = -51.35 (c 1.0, CHCl3). 1H-NMR (CDCl3, 300 MHz) δ 0.83 (t, J = 7.3 Hz, 6H), 1.24-1.50 (m, 4H), 2.29-2.46 (m, 4H), 2.29-2.46 (m, 2H), 3.90 (sa, 1H), 3.92-4.00 (m, 1H), 4.00-4.11 (m, 1H), 5.02 (d, J = 12.3 Hz, 1H), 5.07 (d, J = 12.3 Hz, 1H) 5.58 (d , J = 10.1 Hz, 1H), 7.25.7.46 (m, 8H), 7.50-7.54 (m, 2H). 13C-NMR (CDCl3, 75.5 MHz) δ 11.6, 20.1, 55.8, 56.9, 57.1 (t, JCF = 29.6 Hz), 62.9, 66.9, 121.5 (t, JCF = 249.0 Hz), 125.8 (t, JCF = 6.4 Hz ), 127.8, 128.0, 128.2, 128.4, 130.0, 134.8 (t, JCF = 26.0 Hz), 136.3, 156.3. 19F-NMR (CDCl3,

282.4 MHz) δ -104.8 (dd, JFF = 2489.9 Hz, JHF = 15.1 Hz, 1F), -103.2 (dd, JFF = 248.9 Hz, JHF = 12.2 Hz, 1F). EMAR (FAB) calculated for C24H33F2N2O3 [M + H +]: 435.2471, found: 435.2467.

(2R, 3R) -1-Phenyl-1,1-di-fluoro-3-hydroxy-4- (benzylamino) butane-2-carbamate benzyl (28b). To a solution of epoxide 12b (20 mg, 0.06 mmol) in anhydrous isopropanol (0.5 mL) at room temperature, benzylamine (0.15 mmol) was added and the mixture was heated at 70 ° C with constant stirring, until it was observed the disappearance of the starting epoxide by CCF. Then, the solvent was removed under reduced pressure and the crude obtained was purified by fl ash chromatography, using n-hexane: AcOEt (2: 1) as eluent, the product 28b (17.2 mg, 65%) being isolated as a white solid . Mp = 132-134 ° C (CH2Cl2). [α] D25 = -11.51 (c 1.0, CHCl3). 1H-NMR (CDCl3, 300 MHz) δ 2.14 (sa, 2H), 2.80 (dd, J = 12.6, 6.2 Hz, 1H), 2.88 (dd, J = 12.6, 3.7 Hz, 1H), 3.72 (d, J = 13.3Hz, 1H), 3.77 (d, J = 13.3Hz, 1H), 3.89 (td, J = 6.2, 3.7 Hz, 1H), 4.43-4.58 (m, 1H), 5.00 (d, J = 12.2 Hz , 1H), 5.06 (d, J = 12.2 Hz, 1H), 6.00 (d, J = 9.9 Hz, 1H), 7.24

7.50 (m, 15H). 13C-NMR (CDCl3, 75.5 MHz) δ 50.7, 53.9, 59.7 (t, JCF = 27.0 Hz), 67.1, 67.7, 121.4 (t, JCF = 246.0 Hz),

125.4 (t, JCF = 6.4 Hz), 127.1, 127.9, 128.0, 128.1, 128.4, 128.5, 128.5, 130.2, 134.9 (t, JCF = 25.5 Hz), 136.1, 139.6,

156.5. 19F-NMR (CDCl3, 282.4 MHz) δ -104.9 (d, JFF = 250.5 Hz, 1F), -101.9 (dd, JFF = 250.5 Hz, 1F). EMAR (ESI +) calculated for C25H29N2O3 [M + H +]: 441.2002, found: 441.1989.

Evaluation of the antimicrobial activity of di-fluorobenzyl ethanolamines

An initial evaluation was made by studying the antimicrobial activity of the selected synthetic intermediates. Initially all the compounds in schemes 8,9 and 10 as well as all those included in Table 1 were subjected to different tests to determine their possible application as antimicrobials, against different species of bacteria, gram positive and gram negative, and against fungi, using Agar diffusion technique. This is a semi-quantitative technique that allows to determine the inhibition of microbial growth produced by the compound under study.

Agar diffusion method

The in vitro antimicrobial activity of the compounds was determined against various species (S. aureus ATCC 29213, E. coli ATCC 25922, M. luteus ATCC 49732, M. smegmatis ATCC 19420, and C. albicans ATCC 14053) using a suspension of Frozen cells (0.5 McFarland units) of these microorganisms in Mueller-Hinton culture broth (BBL, BD, Sparks, Md.) with 20% glycerol. After defrosting and homogenizing the previous suspensions, a moistened swab was used to perform a grass seeding on Mueller-Hinton agar plates (BBL, BD, Sparks, Md.). Sterile 6 mm paper discs, BBL, BD, Sparks, Md. Were impregnated with the dissolution of the antimicrobial agents in DMSO and placed on the surface of the agar. The plates were incubated in the air at 37 ° C overnight before reading the inhibition zones.

A similar methodology was used for the evaluation of activities against M. kansasii, N. asteroides, and

N. farcinica. In this case, the suspension used was prepared at 3 McFarland units for the first and 1 McFarland units for Nocardia species. In addition, the medium used for the tests with M. kansasii was Middlebrook and Cohn 7H10 agar (BBL, BD, Sparks, Md.). Plates were incubated at 37 ° C for 7-8 days (M. kansasii) or 2-3 days (Nocardia spp.) Prior to reading the zone of inhibition.

Method of microdilution in broth for Mycobacterium

To the 96-well polystyrene microtiter plates with round bottom (Corning Inc., Corning, NY), 50 µL of 7H10 broth modified per well was added. The antimicrobial agents were prepared at a 4X concentration the maximum concentration tested, and 50 µL of this solution was added to the first well. Subsequently double serial dilutions were made, leaving the last well as a positive control, without antimicrobial agent. To prepare the bacterial isolates, the frozen cultures were thawed and diluted to a final concentration of 1.25 x 105 CFU / mL (working inoculum) in modified 7H10 broth (7H10 agar formulation in which the agar and green were omitted from malachite). The final inoculum was measured by titration and seeding in 7H10 agar with 10% OADC; agar plates were incubated for 1 week at 37 ° C. 50 µL of the work inoculum was added to each well. Microtiter plates were covered with SealPlate sealing adhesive film (Excel Scienti fi c, Wrightwood, CA) and incubated at 37 ° C for 7-8 days prior to reading. The minimum inhibitory concentration (MIC) was defined as the lowest concentration of antimicrobial agent that did not provide visible turbidity. Each bacterial isolate was tested in duplicate.

Broth microdilution method for Nocardia

To the 96-well round-bottom polystyrene microtiter plates (Corning Inc., Corning, NY), 50 μL of modified Mueller-Hinton broth was added per well. The antimicrobial agents were prepared at a 4X concentration the maximum concentration tested, and 50 µL of this solution was added to the first well. Subsequently double serial dilutions were made, leaving the last well as a positive control, without antimicrobial agent. To prepare the bacterial isolates, the frozen cultures were thawed and diluted to a final concentration of 1.25 x 105 CFU / mL (working inoculum) in Mueller-Hinton broth with cationic supplement (following the procedure approved by the Clinical and Laboratory Standards Insitute, CLSI). The final inoculum was measured by titration and seeding on Mueller-Hinton agar. The agar plates were incubated for 4 days at 37 ° C. 50 µL of the work inoculum was added to each well. Microtiter plates were covered with SealPlate sealing adhesive film (Excel Scienti fi c, Wrightwood, CA) and incubated at 37 ° C for 3-4 days prior to reading. The minimum inhibitory concentration (MIC) was defined as the lowest concentration of antimicrobial agent that did not provide visible turbidity. Each bacterial isolate was tested in duplicate.

The intermediate di-fluorobenzyl derivatives of the allylamine and epoxide type studied (4 and 12b, respectively) showed no evidence of inhibition of microbial growth while the di-fluorobenzyl derivative with hydroxyethylamine skeleton 17b showed an antimicrobial activity that proved to be selective against some of the microorganisms tested . Specifically, this compound produced growth inhibition of strains of Mycobacterium smegmatis and Micrococcus luteus but not of other bacteria such as Staphylococcus aureus, Escherichia coli or fungi such as Candida albicans (Table 2, entries 1-5). This selectivity is of special interest since it suggests a mechanism of specific action on special characteristics of some species, as opposed to a nonspecific and generalized mechanism that could be potentially toxic.

The bacteria of the Mycobacterium genus constitute a group of special relevance due to their pathogenicity (they are the cause of tuberculosis, leprosy and other pulmonary, cerebral and cutaneous infections among others) and because of the difficulty in their treatment, due to the differential characteristics of Their bacterial wall confers high resistance to conventional treatments. As a consequence, the search for new active drugs continues to be studied. Therefore, the in vitro evaluation was extended to other Mycobacterium species. It was also extended to Nocardia species, a genus of pathogenic bacteria closely related to Mycobacterium. The results obtained are shown in Table 2, expressed in millimeters, corresponding to the diameter of the growth inhibition halo produced around the disk impregnated in the compound to be tested.

(Table goes to next page) TABLE 2

Compound 17b demonstrated activity against various Mycobacterium species, especially M. smegmatis and M. kansasii. It also produced growth inhibition of N. asteroids and N. farcinica.

Evaluation of in vitro antibacterial activity on Mycobacterium and Nocardia species

Due to the interest of the results obtained in the initial evaluation, it was decided to deepen the study of other structurally related molecules. Thus, a small library of products was prepared to be tested, in order to carry out a structure-activity correlation study (QSAR). The preparation of the rest of diastereoisomers of the compound 17b initially studied and also of other molecules with modifications in various functional groups was discussed.

Ethambutol (EMB) was used as the reference compound in the tests. An ethambutol analogue, SQ-109 developed by Sequella Inc., which is under investigation due to the good results provided as a potential antituberculous agent, was also included in the study in previous studies (Journal of Antimicrobial Chemotherapy 2006, 58, 332-337, Antimicrobial Agents and Chemotheraphy, 2007, 1563-1565). Table 3 shows the results of diffusion in agar, in millimeters of inhibition, of the four diastereoisomers of the structure that had provided the best results initially, together with EMB and SQ-109.

TABLE 3

Agar diffusion assays revealed that the four diastereoisomers were active on various strains of Mycobacteria and Nocardia, especially on N. asteroids 720, strain on which EMB was inactive, which is especially interesting. Few inhibition differences were observed between diastereoisomers, with 17b antibacterial activity slightly better. EMB proved to be much more active on Mycobacterium kansasii than the new compounds, while in the case of Nocardia the differences were insignificant. SQ109 provided the best results on both Mycobacterium and Nocardia.

Other structural modifications that were carried out on the main molecule (compounds, 16a, 16b, 16'a, 16'b, 27a and 28b) following the same process of obtaining that had been carried out with 17 but using the appropriate commercial amines, they led to compounds that turned out to be inactive, a fact that allowed to deduce that both the Cbz group on the nitrogen atom and the presence of the iodine atom on the aromatic ring are of special importance in the biological activity. Preliminary tests of other derivatives with different aromatic substitutions on the amine, subsequently prepared, (compounds 18a, 19a, 18b, 19b, 20b, 21b, 22b, 23a, 24a, 25b, 23b, 26b and 24b), also obtained from the above They seemed to suggest that the most favorable combination for antimicrobial activity was that of product 17b, the rest of the analogs being less active than this or even completely inactive.

In view of the results, we proceeded to analyze the data with a technique that allowed us to determine the bacteriostatic activity quantitatively and in a liquid medium, given that sometimes the agar diffusion technique can give false negatives or inaccurate data due to diffusion problems of the molecules in the agar. Therefore, the method of determining the minimum inhibitory concentration (MIC) by microdilution in culture broth was used. The MIC data (expressed in μg / ml) obtained for the four diastereoisomers together with those of EMB and SQ-109 are shown in Table 4.

TABLE 4

It should be noted that while on Mycobacterium kansasii the MIC of EMB is much lower than that of the new derivatives and even better than that of SQ-109, when it comes to Nocardia, the ethanolamine type derivatives are more potent than EMB (this is relatively inactive ), presenting lower CMI, and very similar in activity to SQ-109, being in this case 17b and 17'b almost as active as SQ-109, with little significant CMI differences. Therefore, it can be concluded that fluorinated ethanolamine-type compounds (especially 17b and 17b) are more active against Nocardia than EMB with MICs similar to those of SQ-109.

Di-fluorobenzyl ethanolamines according to the invention can be used in the preparation of a medicament for the treatment of leprosy, tuberculosis or some conditions related to any of these diseases. For this, a pharmaceutical composition characterized in that it comprises at least one stereoisomer of one or more di-fluorobenzyl ethanolamines or a pharmaceutically acceptable salt thereof, together with one or more vehicles and / or diluents necessary to be administered orally, transdermally, can be made. parental or inhalative. In such cases, di-fluorobenzyl ethanolamines are presented as active constituents in the appropriate presentation forms for the different forms of administration such as, for example, tablets, capsules, suppositories, solutions, juices, emulsions or dispersible powders. The tablets may also be delayed release.

References

1 Spigetman M.K., The Journal of Infectious Diseases, 2007, 196, S28-34.

2Sacchettini, J.c. Rubin, E.J., Freudlinch, J.S., Microbiology 2008, 6, 41-52.

3Chatterjee, D., Biopolymers, 1997, 1, 579-588.

4 Gutierrez-Lugo, M.T., Bewley, C.A., J. Med. Chem. 2008, 51, 2606-2612.

5Ralp, A.P., Anstey, N.M., Kelly, P.M., Clinical Infectious Diseases, 2009, 49, 574-583.

6Kuduk, S.D., Marco, C.N.D., Pitzenberger, S.M., Tsou, N., Tetranhedron Lett., 2006, 47, 2377-2381.

7 Philippe, Christine et al. "Synthesis of New Trii fl uoromethylated Hydroxyethylamine-Based Scaffolds" Eur. J.

Org. Chem. 2009, 5215-5223.

8 Two Saints, Mickael et al., Tetrahedron Letters, 2009, 50, 857-859.

9Purser, S., Moore, P.R .; Swallow, S .; Governeur, V., Chem. Soc. Rev. 2008, 37, 320-330; Muller, K .; Faeh, C .;

Diederich, F., Science, 2007, 317, 1881-1886; Hagmann, W.K., J. Med. Chem. 2008, 51, 4359-4369.

10Khetan, S.K .; Collins, T.J. Chem. Rev. 2007, 107, 2319-2364.

11 Scott E. Denmark et al., J. Org. Chem. 1995, 60, 1391-1407.

12R. Smits, C.D. Cadicamo, K. Burger, B. Koksch, Chemical Society Reviews 2008, 37, 1727-1739.

13S. Fustero, J.F. Sanz-Cerverea, J.L. Aceña, M. Sánchez-Roselló, Synlett 2009, 525-549.

14 Sani, M .; Volontery, A .; Zanda, M., ChemMedChem. 2007, 2, 1693-1700; Pesenti, C .; Arnone, A .; Bellosta,

S .; Bravo, P .; Canavesi, M .; Corradi, E .; Frigerio, M .; Meille, S.V .; Monetti, M .; Panzeri, W.; Viani. F.; Venturini, R .; Zanda, M., Tetrahendron, 2001, 57, 6511-6522.

15Fustero, S .; Flores, S .; Cuñat, A.C .; Jiménez, D .; From the well; C., Sanz-Cervera, J.F., Journal of Fluorine Chemistry 2007, 128, 1248-1254.

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18 Although a representative sample was conveniently separated for the characterization of both isomers.

19 Peltier, H.M .; McMahon, J.P .; Patterson, A.W .; Ellman, J.A. J. Am. Chem. Soc. 2006, 128, 16018-16019.

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Claims (23)

1. Di-fluorobenzyl ethanolamines represented by the general formula (I), where R is a protective group and Ar is a derivative of benzene, furan, thiophene or pyridine. Where: a) the protecting group R may be benzyloxycarbonyl, tert-butyloxycarbonyl; Y b) Ar are derivatives of benzene, furan, thiophene or pyridine, which may independently possess fluorine, chlorine, bromine, iodine, methoxy as substituents.

2.

Any di-fluorobenzyl ethanolamine (I) according to claim 1, characterized in that it is composed of a mixture of different isomers.

3.

Any di-fluorobenzyl ethanolamine (I) according to claim 1, characterized in that it is composed of a single isomer.

Four.

Di-fluorobenzyl ethanolamine according to claims 1-3, characterized in that the protecting group is tert-butyl carbamate and Ar is 3-iodophenyl and is selected from (2R, 3S) -1-Phenyl-1,1-di-fluoro-3 tert-butyl hydroxy4- (3-iodobenzylamino) butanyl-2-carbamate (16a); 16b: tert-butyl (16-phenyl-1,1-di-fluoro-3-hydroxy-4- (3-iodobenzylamino) butanyl-2-carbamate (16b):

(2S, 3R) -1-Phenyl-1,1-di-fluoro-3-hydroxy-4- (3-iodobenzylamino) tert-butyl butanyl-2-carbamate (16'a) or (2S, 3S) 1-Phenyl -1,1-di-fluoro-3-hydroxy-4- (3-iodobenzylamino) tert-butyl butanyl-2-carbamate (16'b): 5. Di-fluorobenzyl ethanolamine according to claims 1-3, characterized in that the protecting group is benzyl carbamate and Ar is 3-iodophenyl and is selected from (2R, 3S) -1-Phenyl-1,1-di-fluoro-3 benzyl hydroxy-4- (3-iodobenzylamino) butane-2-carbamate (17a); (2R, 3R) -1-Phenyl-1,1-di-fluoro-3-hydroxy-4- (3-iodobenzylamino) butaneyl-2-carbamate benzyl (17b): (2S, 3R) -1-Phenyl-1,1-di-fluoro-3-hydroxy-4- (3-iodobenzylamino) bucynyl-2-carbamate of benzyl (17'a) or (2S, 3S) -1Phenyl-1, Benzyl-di-fluoro-3-hydroxy-4- (3-iodobenzylamino) butane-2-carbamate (17'b).

6.

Di-fluorobenzyl ethanolamine according to claims 1-3, characterized in that the protecting group is benzyl carbamate and Ar is 3-fl uorophenyl and is selected from (2R, 3S) -N- {2- [1-Phenyl-1,1 di-fluoro-4- (3-fluorophenylmethyl) amino-3-hydroxy]} benzyl carbamate (18a) or (18b): (2R, 3R) -N- {2- [1-Phenyl-1,1-di-fluoro-4- Benzyl (3-fl uoro-phenylmethyl) amino-3-hydroxy] carbamate (18b):

7.

Di-fluorobenzyl ethanolamine according to claims 1-3, characterized in that the protecting group is benzyl carbamate and Ar is 3-chlorophenyl and is selected from (2R, 3S) -N- {2- [4- (3-Chlorophenylmethyl) benzyl (19a) or (2R, 3R) -N- {2- [4 (3-Chlorophenylmethyl) amino-1-phenyl-1,1- amino-1-phenyl1,1-di-fluoro-3-hydroxy]} carbamate benzyl di-fluoro3-hydroxy]} carbamate (19b):

8.

Di-fluorobenzyl ethanolamine according to claims 1-3, characterized in that the protecting group is benzyl carbamate and Ar is 3-methoxyphenyl, such as (2R, 3R) -N- {2- [1-Phenyl-1, 1-di-fluoro-3-hydroxy4- (3-methoxyphenylmethyl) amino]} benzyl carbamate (20b):

9.

Di-fluorobenzyl ethanolamine according to claims 1-3, characterized in that the protecting group is benzyl carbamate and Ar is 2-thienyl, such as (2R, 3R) -N- {2- [1-Phenyl-1, 1-di-fluoro-3-hydroxy-4- (2-thienylmethyl) amino]} benzyl carbamate (21b).

10.

Di-fluorobenzyl ethanolamine according to claims 1-3, characterized in that the protecting group is benzyl carbamate and Ar is 2-furyl, such as (2R, 3R) -N- {2- [1-Phenyl-1, 1-di-fluoro-4- (2-furylmethyl) amino-3-hydroxy]} benzyl carbamate (22b):

eleven.

Di-fluorobenzyl ethanolamine according to claims 1-3, characterized in that the protecting group is tert-butyl carbamate and Ar is 3-fl uorophenyl, such as (2R, 3S) -N- {2- [1-Phenyl-1 , 1-di-fluoro-4- (3-fluorophenylmethyl) amino-3-hydroxy]} tert-butyl carbamate (23a) or (2R, 3R) -N- {2- [1-Phenyl-1,1-di-fluoro-4 - (3-fl uorophenylmethyl) amino-3-hydroxy]} tert-butyl carbamate (23b).

12.

Di-fluorobenzyl ethanolamine according to claims 1-3, characterized in that the protecting group is tert-butyl carbamate and Ar is 2-thienyl, such as (2R, 3S) -N- {2- [1-Phenyl- 1,1-di-fluoro-3-hydroxy4- (2-thienylmethyl) amino]} tert-butyl carbamate (24a) or (2R, 3R) -N- {2- [1-Phenyl-1,1-di-fluoro-3 tert-butyl hydroxy-4- (2-thienylmethyl) amino]} carbamate (24b):

13.

Di-fluorobenzyl ethanolamine according to claims 1-3, characterized in that the protecting group is tert-butyl carbamate and Ar is 3-chlorophenyl, such as (2R, 3R) -N- {2- [4- (3 - Chlorophenylmethyl) amino1-phenyl-1,1-di-fluoro-3-hydroxy]} tert-butyl carbamate (25b):

14.

Di-fluorobenzyl ethanolamine according to claims 1-3, characterized in that the protecting group is tert-butyl carbamate and Ar is 3-methoxyphenyl, such as (2R, 3R) -N- {2- [1-Phenyl- Tert-butyl 1,1-di-fluoro-3-hydroxy-4- (3-methoxyphenylmethyl) amino]} carbamate (26b):

fifteen.

Di-fluorobenzyl ethanolamine according to claims 1-3, characterized in that the protecting group is benzyl carbamate and Ar is phenyl, such as (2R, 3R) 1-Phenyl-1,1-di-fluoro-3-hydroxy-4 - benzyl (benzylamino) butanyl-2-carbamate (28b)

16. Process for obtaining di-fluorobenzyl ethanolamines (I) or a pharmaceutically acceptable salt thereof according to claims 1-15, characterized in that it is obtained from di-fluorobenzyl allylamines 4, or any of its isomers, by the following steps : a) Introduction of the protective group R by reaction of the fluorobenzyl allylamine 4 with the compound containing the desired protecting group, for example with benzyloxycarbonyl to obtain carbamate 10: b) Formation of intermediate epoxides 12a and 12b by reaction of di-fluorobenzyl allylamines 10 for example with tri-fluoromethyldioxyran: c) Reaction of epoxides 12a and 12b with the desired compound containing the Ar group that is desired to be introduced, for example, with 3-iodobenzylamine to form di-fluorobenzyl ethanolamine 17a:

17.

Process for obtaining di-fluorobenzyl ethanolamines (I) or a pharmaceutically acceptable salt thereof, according to claim 16, characterized in that the isomers of the epoxides obtained in step (b) are separated, the reaction of step (c) with each isomer independently.

18.

Process for obtaining di-fluorobenzyl ethanolamines (I) or a pharmaceutically acceptable salt thereof, according to claim 16, characterized in that the isomers of the epoxides obtained in step (b) are separated by column chromatography with silica gel. .

19.

Use of di-fluorobenzyl ethanolamines (I) or a pharmaceutically acceptable salt thereof, according to claims 1-15, for the preparation of a medicament for antimicrobial treatment.

twenty.

Use of a di-fluorobenzyl ethanolamine (I) or one of its isomers or a pharmaceutically acceptable salt thereof, according to claim 19, for the preparation of a medicament for the treatment of tuberculosis.

twenty-one.

Use of a di-fluorobenzyl ethanolamine (I) or one of its isomers or a pharmaceutically acceptable salt thereof, according to claims 19 and 20, for the preparation of a medicament for the treatment of leprosy.

22

Pharmaceutical composition according to claims 1 to 15, characterized in that it comprises at least one stereoisomer of one or more di-fluorobenzyl ethanolamines or one of its pharmaceutically acceptable salts, according to claims 1 to 19, together with one or more vehicles and / or diluents necessary for use according to claims 19-21.

SPANISH OFFICE OF THE PATENTS AND BRAND Application no .: 201000997 SPAIN Date of submission of the application: 30.07.2010 Priority Date: REPORT ON THE STATE OF THE TECHNIQUE 51 Int. Cl.: See Additional Sheet RELEVANT DOCUMENTS

Category

Documents cited Claims Affected

TO

PHILIPPE, C. et al. "Synthesis of New Trifluoromethylated Hydroxyethylamine-Based Scaffolds." European Journal of Organic Chemistry 2009, pages 5215-5223. [Available online on 03.09.2009]. See page 5215, summary and introduction; page 5216, table 1, compound 2d. 1-22

TO

US 20070117793 A1 (GHOSH, A.K. et al.) 24.05.2007, paragraphs [0005] - [0006], compound I; paragraph [0158], scheme 3; paragraphs [0108], [0114], [0115], [0118]; page 21, example 1.1. 1-22

TO

DOS SANTOS, M. et al. "Improved Ritter reaction with CF3-containing oxirane for an access to central units of protease inhibitors." Tetrahedron Letters 2009, Volume 50, pages 857-859. [Available online on 02.10.2008]. See page 857, summary; page 858, scheme 3; page 859, scheme 4, compounds 3b-j. 1-22

TO

FUSTERO, S. et al. "Synthesis of fluorinated allylic amines: Reaction of 2- (trimethylsilyl) ethyl sulfones and sulfoxides with fluorinated imines." Journal of Fluorine Chemistry 2007, Volume 128, pages 1248-1254. See page 1248, summary; page 1249, scheme 3. 1-22

TO

WO 2004050519 A1 (GLAXO GROUP LIMITED) 06.17.2004, page 10, compound III; page 13, scheme; pages 2, lines 32-34. 1-22

Category of the documents cited X: of particular relevance Y: of particular relevance combined with other / s of the same category A: reflects the state of the art O: refers to unwritten disclosure P: published between the priority date and the date of priority submission of the application E: previous document, but published after the date of submission of the application

This report has been prepared • for all claims • for claims no:

Date of realization of the report 04.10.2011

Examiner G. Esteban García Page 1/4

REPORT OF THE STATE OF THE TECHNIQUE Application number: 201000997 CLASSIFICATION OBJECT OF THE APPLICATION  C07C215 / 28 (2006.01) C07C213 / 04 (2006.01) A61K31 / 137 (2006.01) A61P31 / 04 (2006.01) A61P31 / 06 (2006.01) A61P31 / 08 (2006.01) Minimum documentation sought (classification system followed by classification symbols) C07C, A61K, A61P Electronic databases consulted during the search (name of the database and, if possible, search terms used) INVENES, EPODOC, WPI, REGISTRY, CAPLUS, EMBASE, BIOSIS, MEDLINE, XPESP, NPL, PUBMED State of the Art Report Page 2/4  WRITTEN OPINION Application number: 201000997 Date of Completion of Written Opinion: 04.10.2011 Statement

Novelty (Art. 6.1 LP 11/1986)

Claims 1-22 Claims IF NOT

Inventive activity (Art. 8.1 LP11 / 1986)

Claims 1-22 Claims IF NOT

The application is considered to comply with the industrial application requirement. This requirement was evaluated during the formal and technical examination phase of the application (Article 31.2 Law 11/1986).  Opinion Base.- This opinion has been made on the basis of the patent application as published. State of the Art Report Page 3/4  WRITTEN OPINION Application number: 201000997 1. Documents considered.- The documents belonging to the state of the art taken into consideration for the realization of this opinion are listed below.

Document

Publication or Identification Number publication date

D01

PHILIPPE, C. et al. European Journal of Organic Chemistry 2009, pp. 5215-5223 03.09.2009

D02

US 20070117793 A1 05/24/2007

D03

DOS SANTOS, M. et al. Tetrahedron Letters 2009, Vol. 50, pp. 857-859 02.10.2008

2. Statement motivated according to articles 29.6 and 29.7 of the Regulations for the execution of Law 11/1986, of March 20, on Patents on novelty and inventive activity; quotes and explanations in support of this statement The object of the invention is a series of difluorobenzyl ethanolamines of the general formula (I), a multistage process for obtaining them, their use for the preparation of a medicament and a pharmaceutical composition comprising at least one compound of formula (I). Document D01 discloses peptidomimetics derived from trifluoromethyl-hydroxyethylamine that are prepared by regioselective opening of an epoxide with compounds that comprise an amine group, such as amino acids and dipeptides, and that have activity as protease inhibitors (see page 5215, summary and introduction ). Specifically, the 2d derivative, obtained from epoxy 1a, which differs from the compound of the invention is disclosed in that it comprises a CF3-moiety instead of the -CF2Ph present in the latter (see page 5216, table 1). Document D02 discloses a series of compounds of formula (I) that comprise an ethanolamine group and that have activity as 1-secretase inhibitors and are useful for the treatment of Alzheimer's disease (see paragraphs [0005] - [0006], compound I). These compounds are prepared by a multi-stage process that runs through an intermediate 9s (see paragraph [0158], scheme 3), whose general formula includes the compound (I) of the invention, which would result from the selection of substituents suitable within all the possibilities listed for groups A2, L2, L3, R5 and R7 in document D02 (see paragraphs [0108], [0114], [0115], [0118]). Therefore, it is considered that the compound of the invention is new, and that the determination of the substituents mentioned among all the alternatives included in document D02 constitutes a multiple selection that requires the exercise of inventive activity. On the other hand, this document discloses t-butyl 3-hydroxy-4- (3-methoxybenzylamino) -1-phenylbutan-2-yl-carbamate, and a process for its preparation by opening the epoxide present in the ( T-Butyl 1-oxyranyl-2-phenylethyl) carbamate with 3-methoxybenzylamine (see page 21, example 1.1). This compound differs from the compound of the invention in that the benzyl group is not substituted, that is, it lacks the two fluorine atoms. Document D03 discloses a series of amino alcohols of formula 3 that are obtained by opening the corresponding epoxides by a reaction of Ritter with nitriles (see page 857, summary; page 858, scheme 3), and which have activity as protease inhibitors, due to the presence of fluoroalkyl groups, which makes them mimetics of natural hydrophobic residues, such as benzyl or isopropyl. These compounds differ from those of the invention in that it comprises a CF3 moiety instead of the CF2Ph present in the latter, in addition to having the second amine group of the amide-shaped molecule (see page 859, scheme 4, compounds 3b-j ). The cited documents show only the state of the art of the field to which the invention belongs. None of them, taken alone or in combination with the others, disclose or contain any suggestion that could direct the person skilled in the art to difluorobenzyl ethanolamines of the general formula (I) (independent claim 1) and, therefore, also not to a procedure in several stages for obtaining them (independent claim 16), their use for the preparation of a medicament (independent claim 19) and a pharmaceutical composition comprising at least one compound of formula (I) (independent claim 22). Therefore, it is considered that the object of the invention meets the requirements of novelty and inventive activity set forth in Articles 6.1 and 8.1 of the Patent Law. State of the Art Report Page 4/4